LOCUS       Exported                7764 bp ds-DNA     circular SYN 10-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAJ30
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7764)
  AUTHORS   Joers lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Nov 10, 2020 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7764
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=pBR322 ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    775..917
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1019..1210)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2019..3101)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3102..3179)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        3488..3506
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3507..3531
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             3575..4807
                     /codon_start=1
                     /label=6His-KshA5
                     /translation="MGSSHHHHHHSSGLVPRGSHMSIDTARSGSDDDVEIREIQAAAAP
                     TRFARGWHCLGLLRDFQDGKPHSIEAFGTKLVVFADSKGQLNVLDAYCRHMGGDLSRGE
                     VKGDSIACPFHDWRWNGKGKCTDIPYARRVPPIAKTRAWTTLERNGQLYVWNDPQGNPP
                     PEDVTIPEIAGYGTDEWTDWSWKSLRIKGSHCREIVDNVVDMAHFFYIHYSFPRYFKNV
                     FEGHTATQYMHSTGREDVISGTNYDDPNAELRSEATYFGPSYMIDWLESDANGQTIETI
                     LINCHYPVSNNEFVLQYGAIVKKLPGVSDEIAAGMAEQFAEGVQLGFEQDVEIWKNKAP
                     IDNPLLSEEDGPVYQLRRWYQQFYVDVEDITEDMTKRFEFEIDTTRAVASWQKEVAENL
                     AKQAEGSTATP"
     CDS             3587..3604
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4884..5999
                     /codon_start=1
                     /label=6His-KshB
                     /translation="MGSSHHHHHHSSGLVPRGSHMTTVEVPHGSRSVILTVSAVVEETA
                     DTRSIVFAVPDELRDKFAYRPGQFLTLRIPSDRTGSVARCYSLASSPFTDDAPKVTVKR
                     TSDGYGSNWLCDNIATGQTLEVLPPAGVFTPKSLDHDFLLFGAGSGITPVISILKSALT
                     QGGGKVVLVYANRDEKSVIFAEELRALAEKYPTRLTVVHWLESVQGLPTADQLAAIAAP
                     YESYEAFMCGPGPFMDTVHQALNTVGMPRARVHAEVFNSLSGDPFADQAPVEVSDEDAA
                     DAATVEVELDGEVHKLSWPRKQTLVDIMLAKGIDVPYSCQEGECGSCACTVLEGKVEME
                     NCDVLDPEDIEAGYILGCQARPVTDHLKIEF"
     CDS             4896..4913
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6063..6110
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6147..6602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6629..6733
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6734..7594
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc
      841 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
      901 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      961 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     1021 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     1081 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     1141 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     1201 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     1261 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     1321 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     1381 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     1441 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     1501 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     1561 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     1621 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     1681 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     1741 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     1801 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     1861 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     1921 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     1981 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     2041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2101 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     2161 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     2221 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     2281 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     2341 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     2401 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     2461 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     2521 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     2581 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     2641 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     2701 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     2761 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     2821 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     2881 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     2941 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     3001 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     3061 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     3121 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     3181 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     3241 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     3301 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     3361 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     3421 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     3481 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
     3541 ataattttgt ttaactttaa gaaggaggta taccatggga tctagtcacc accaccatca
     3601 tcattcctca ggactggtcc cccgcggttc gcacatgtcc atcgacaccg cccgctcagg
     3661 gtctgacgat gacgttgaaa ttcgtgaaat tcaggccgcc gcggccccta cacgttttgc
     3721 tcgtggctgg cattgccttg gattgttacg cgattttcag gatggaaagc cacacagcat
     3781 cgaggctttc ggcacaaaac ttgtagtctt tgccgactcg aagggccaac ttaacgtctt
     3841 ggacgcctac tgtcgtcaca tgggcggaga cctttcacgt ggtgaagtaa aaggtgatag
     3901 catcgcttgc ccgtttcacg attggcgctg gaatggcaag ggtaagtgca cggacattcc
     3961 gtatgctcgc cgtgttcccc cgatcgccaa aacccgtgcc tggactacct tagagcgtaa
     4021 cggacagttg tacgtatgga atgatcctca gggtaacccg ccccccgagg acgttacaat
     4081 tcctgaaatc gcgggatatg gaaccgacga gtggacagat tggtcatgga aaagtcttcg
     4141 tattaaagga tcgcactgtc gcgaaatcgt agacaatgtc gtcgacatgg cacacttttt
     4201 ctatattcat tattcgttcc ctcgttactt caagaatgtt tttgagggtc acacggcgac
     4261 acaatatatg cactcaacag gccgtgaaga cgtgatcagt ggcactaact acgatgatcc
     4321 taacgcagag cttcgcagtg aggccactta cttcggtcca agctacatga tcgattggtt
     4381 agaatctgac gctaatggcc agacgatcga gactatctta atcaactgcc actaccccgt
     4441 atcgaataat gagttcgtcc tgcagtatgg ggcgattgta aagaagttgc caggggtgag
     4501 cgatgaaatc gcagctggta tggctgagca atttgccgag ggcgtgcaac ttggcttcga
     4561 gcaagatgtg gagatctgga agaataaggc tccaatcgat aatccactgc tgtcggagga
     4621 ggacggtcca gtctaccaat tgcgtcgttg gtaccaacag ttttatgtgg atgtcgagga
     4681 catcactgag gacatgacta aacgtttcga gttcgagatt gatacgactc gcgccgttgc
     4741 gagttggcag aaggaagtgg ccgaaaattt ggctaaacag gcggaagggt caaccgctac
     4801 cccatagagc atctgcagaa ttccagcaca ctggcggccg ttactagaaa taattttgtt
     4861 taactttaag aaggaggtat accatggggt ctagtcacca ccatcatcat cattcatccg
     4921 ggttagtgcc acgtggctct cacatgacaa cggtcgaagt accgcatggc tcgcgcagcg
     4981 taatcctgac agtttcagcc gtggtggaag agaccgccga cacccgttca attgttttcg
     5041 ctgttccaga tgaactgcgc gacaaattcg cgtatcgccc tggccagttt ctgacgctgc
     5101 gtatcccatc cgatcgcacc ggatctgttg ctcgttgtta ttcgttagcg agctctccgt
     5161 ttacggacga tgccccaaag gtgactgtaa agcgcacctc ggacggctac ggatctaatt
     5221 ggttgtgcga caatattgcg acaggccaaa ccctggaagt gcttcctccc gcaggtgtat
     5281 ttacacccaa gtccttggac cacgattttt tgttatttgg ggcggggtcg gggattaccc
     5341 ctgtgatttc catcttaaag tccgcattaa cccagggcgg cggaaaagtc gttcttgtgt
     5401 atgcaaaccg tgatgagaag tctgtgatct ttgcggaaga attacgcgct ctggcggaga
     5461 agtacccgac ccgcctgacc gtggtgcact ggcttgagag cgtacaaggg ttaccaaccg
     5521 ccgatcaatt ggccgcgatt gctgcacctt atgaatcgta tgaagccttc atgtgtggcc
     5581 cagggccttt tatggatacc gtgcatcaag ccttgaatac agttggaatg cctcgtgcgc
     5641 gcgtgcatgc tgaggtattc aattctttaa gcggagaccc ttttgcagac caagcaccgg
     5701 tagaggtcag cgacgaggac gccgctgacg ccgcgactgt cgaggtcgaa ttagatggcg
     5761 aggttcataa gttatcctgg ccgcgtaagc agaccttggt cgatattatg ttagcgaagg
     5821 ggattgacgt gccgtactcc tgtcaagagg gcgaatgtgg aagttgtgca tgtactgtac
     5881 ttgagggaaa agttgaaatg gagaattgtg acgtattaga tccggaagac attgaggccg
     5941 ggtacatcct tggatgtcaa gcgcgccctg ttacagacca tctgaaaatc gagttctgag
     6001 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     6061 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     6121 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     6181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     6241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     6301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     6361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     6421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     6481 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     6541 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     6601 ttcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6661 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6721 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6781 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6841 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6901 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6961 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     7021 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     7081 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     7141 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     7201 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7261 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7321 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7381 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7441 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7501 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7561 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7621 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7681 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7741 cgtagaaaag atcaaaggat cttc
//
