LOCUS       sequence-482805-        9634 bp    DNA     circular SYN 11-AUG-2025
DEFINITION  Phagemid for SC101 and sfGFP. Contains p15A, CmR, pac site, AraC, 
            C1, and pBAD..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9634)
  AUTHORS   Al'Abri IS, Haller DJ, Li Z, Crook N
  TITLE     Inducible directed evolution of complex phenotypes in bacteria.
  JOURNAL   Nucleic Acids Res. 2022 Jun 10;50(10):e58. doi: 10.1093/nar/gkac094.
  PUBMED    35150576
REFERENCE   2  (bases 1 to 9634)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2022-06-10"; volume: "50(10):e58. doi"; pages: "
            10.1093/nar/gkac094"
COMMENT     Sequence Label: ISA410: sfGFP
FEATURES             Location/Qualifiers
     source          1..9634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..223
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             271..1221
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(1969..1996)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2012..2083)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2095..2808)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     RBS             2815..2826
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      complement(6267..6294)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6310..6381)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(6633..6917)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(6691..6710)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             6944..7822
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSRLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      7884..7927
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     protein_bind    complement(7951..7975)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             8096..8755
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(8162..8181)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      8776..8870
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gagttataca cagggctggg atctattctt tttatctttt tttattcttt ctttattcta
       61 taaattataa ccacttgaat ataaacaaaa aaaacacaca aaggtctagc ggaatttaca
      121 gagggtctag cagaatttac aagttttcca gcaaaggtct agcagaattt acagataccc
      181 acaactcaaa ggaaaaggac tagtaattat cattgactag cccatctcaa ttggtatagt
      241 gattaaaatc acctagacca attgagatgt atgtctgaat tagttgtttt caaagcaaat
      301 gaactagcga ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat tttatgctgt
      361 gtggcactac tcaaccccac gattgaaaac cctacaagga aagaacggac ggtatcgttc
      421 acttataacc aatacgctca gatgatgaac atcagtaggg aaaatgctta tggtgtatta
      481 gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc tttggttaaa
      541 ggctttgaga ttttccagtg gacaaactat gccaagttct caagcgaaaa attagaatta
      601 gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat aaaatataat
      661 ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga gtggttatta
      721 aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct tgatgaattt
      781 aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa ccaatgggtt
      841 ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt ggttgataag
      901 cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca aatggatctc
      961 gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc aacaaccatt
     1021 acatcagatt cctacctaca taacggacta agaaaaacac tacacgatgc tttaactgca
     1081 aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag taagtatgat
     1141 ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga gaacatactg
     1201 gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg aagcatcaag
     1261 actaacaaac aaaagtagaa caactgttca ccgttacata tcaaagggaa aactgtccat
     1321 atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac gagtttttgg
     1381 tgcattcaaa gctgttcacc atgaacagat cgacaatgta acagatgaac agcatgtaac
     1441 acctaataga acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa ttgaacacct
     1501 gagacaactt gttacagctc aacagtcaca catagacagc ctgaaacagg cgatgctgct
     1561 tatcgaatca aagctgccga caacacggga gccagtgacg cctcccgtgg ggaaaaaatc
     1621 atggcaattc tggaagaaat agcgctttca gccggcaaac cggctgaagc cggatctgcg
     1681 attctgataa caaactagca acaccagaac agcccgtttg cgggcagcaa aacccgtacc
     1741 ctaggtctag ggcggcggat ttgtcctact caggagggtt cgtaacatct ctgtaactgc
     1801 ttgagacgtg gactgttgtg atatgccata tatgcagtgg tgttcgccgg cgctggttcc
     1861 ggctgtcttg cttagttgtg agtcgccagg cagtcatcgg tatgatctgt acatgattcg
     1921 tcagtgtgag caccactgac gagcagattt ccagctgaga cctataaacg cagaaaggcc
     1981 cacccgaagg tgagccagtg tgactctagt agagagcgtt caccgacaaa caacagataa
     2041 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gtcatttgta
     2101 cagttcatcc ataccatgcg tgatgcccgc tgcggttacg aactccagca gaaccatatg
     2161 atcgcgtttc tcgttcggat ctttagacag aacgctttgc gtgctcagat agtgattgtc
     2221 tggcagcaga acaggaccat caccgattgg agtgttttgc tggtagtgat cagccagctg
     2281 cacgctgcca tcctccacgt tgtggcgaat tttaaaattc gctttaatgc catttttttg
     2341 tttatcggcg gtgatgtaaa cattgtggct gttaaaattg tattccagct tatggcccag
     2401 gatattgccg tcctctttaa agtcaatgcc tttcagctca atgcggttta ccagggtatc
     2461 gccttcaaat ttcacttccg cacgcgtttt gtacgtgccg tcatccttaa aggaaatcgt
     2521 gcgttcctgc acatagcctt ccggcatggc ggacttgaag aagtcatgct gcttcatatg
     2581 gtccggataa cgagcaaagc actgaacacc ataagtcagc gtcgttacca gagtcggcca
     2641 aggaaccggc agtttaccag tagtacagat gaacttcagc gtcagtttac cattagttgc
     2701 gtcaccttca ccctcgccac gcacggaaaa cttatgaccg ttgacatcac catccagttc
     2761 caccagaata gggacgacac cagtgaacag ctcttcgcct ttacgcatct agtatttctc
     2821 ctctttctct agtagctaag atccgaacct gccgtttctt gagttgccgc gatgttaaga
     2881 aaacattcat aaattaaatg tgaattgccg cacacattat taaataagat ttacaaaatg
     2941 ttcaaaatga cgcatgaaat cacgtttcac tttcggtctc acgttgtatt gcgacgaatt
     3001 gccacgttgt cgcagtgtct gtactgtggg ttaatgtctg tgatcgccgt cgtgagttcg
     3061 tgtcatccgc gcatgattca gatggtcctg gagatcgttg cttgtaccgg ctgtttgtgg
     3121 ctctaccacg tctcatcaga gatccgaacg gtttattaca tacatcaggt aaaactgacc
     3181 gataagccgc tttcttttgg gtatagtgtc gtggacagtc attcatcttt ctgcccctcc
     3241 aaaagtaaaa acccgccgaa gcgggttttt acgtaaaaca ggtgaaactg accgataagc
     3301 cgctttcttt tgggtatagt gtcgtggaca gtcattcatc tttctgcccc tccaaaagca
     3361 aaaacccgcc gaagcgggtt tttacgtaaa ccaggtgaaa ctgaccgata agccgctttc
     3421 ttttgggtat agcgtcgtgg acagtcattc atctttctgc ccctccaaaa gcaaaaaccc
     3481 gccgaagcgg gtttttacgt aaatcaggtg aaactgaccg ataagccggg ttctgtcgtg
     3541 gacagtcatt catctaggcc agcaatcgcc agatccttaa tcatcatccc cgtacagttt
     3601 gcgggtaact tcttcgtctt cctcctcgtc ttcgtccaga tcctgttcaa gccatgcttc
     3661 gtttgatatg ccttccgcat cgacatcacc ataaccacct gtatcgacga tatcggcaat
     3721 ttcttctcta cgatgctcaa tccacaatgc ggcatcagcg cggcggctgg cggcccgttc
     3781 tcgcgtgatt ttatccagat cctcaagtga tggagcgcca gatgctgttt ggttttcctc
     3841 atcatcggta ttggtcttag gagcacgcag atcggctttg atttgctcca gcatcagggg
     3901 cggcactttt cctccatgcg cctcgatgaa ctcagccgct tccagcactg accagttatt
     3961 ttcacgcttt cgttcgtatg ccagcttaac aatgccagct tgccccatag ataaagcgtg
     4021 cttttccgcc tcccggcttt cttttcgata gttattccgg atgctgtaaa tggtgttgat
     4081 caggctgctt atctgcgcgg aacagctgtt tagcatgctc gcgatacggt attcaggcgg
     4141 agtaccttca tcatcgtctt tttgctgatc gcgcatttcc tgcaccaggc gaatacacgt
     4201 atccctggcg ttctccagca taaggagatg agaaagagac ttttccagaa gagtggtttc
     4261 cagaacatcg gccccggacc gacgcaacat agcgcgcgcg gccttccgcg cttcaacgtt
     4321 atctatcagg taatcgccaa cttcgaactc aaagcgttca ccatcatcat ccagggtgtc
     4381 gcgctccagg cgatcacgta aggtccggtg ggcgcgggtg atcacgtcat gatcatctga
     4441 acgatcattt atgcgcttat tttggcgctt cgcattctcg actgcggcac tgacaacggc
     4501 attaactctt tgtttttccg ctatttcagc cgcaatgtga tcacctgcat gttgatcatt
     4561 agagtgatca atgatcatgc tttttagtgg cttcctgact ggcttatttg gcttgcggct
     4621 gtccgtagtc ctggtgtctt ctttgaaggc acggagataa cgacgtgcgg tattagggtt
     4681 aagattaaac tcggcggcat actgtgcgat ggtgtaacca ccatctcgcg ccaggcgagc
     4741 aaaattcttc ttgtgatcgt cccaggtcac ttatgcttcc tttcgagatc ccaacaatat
     4801 tcctactcaa tgaacaaatg actactcgta gaatcggtta acacaccaga ttcagatcct
     4861 taccctctga atcctgccgg tataccccat tgttcgttat ctttattttt ggctaaaacc
     4921 gcattaagag cttcgtttac cgtcatgcaa tgcggtaggt tatcgaagtt tgatatcccg
     4981 ccaatatcag gcgaacgctt gtttttcagg taagcatatt tccgcgcagc cgcctctact
     5041 ttctgcttga actcatgttt ttgagtgcgt tttttggata accgcagatt gtcagccttt
     5101 gcttttgcct tagcgatcca tgaagtcaat tttttgaggc tggttgttcc ggcaccgccg
     5161 gaaactgatc tttttgtttt tttaacttgt gacttcttat tctttattgc cacgtcatcc
     5221 tgacaggggg agggggtatc attttgacat gggggtgtgg ataaaaaatt aaataaagcc
     5281 aatgtcttag cgagaacagc tttaaccttg gttgccgctg acgaaatctt taatttgctt
     5341 tctatcagcg catttttggc ttgttgtgcg aaggccaaaa aggatggtgt aaaccggtac
     5401 aggttagcgc gacgttcacg gtgatcgccg ataacaatct ctacagacag aatacctttg
     5461 tttacagctt cacggaatgc acgaacgacg gttgattggc tataaccagt ttctgccgcg
     5521 atcaggcggt gaggcttgtg aatgaagtat tcactggttg ttgccgcgag atttgcacat
     5581 tgcgacagga tatgcccggc gctacgggat agaccggagt gtgttacaaa gcaggccaat
     5641 tcatagccag aaaaagtaaa atcgctttaa gatccccgag ttctcttaaa ccaaggttta
     5701 ggattgaaat gatgacgccg gaaacttctt ataaagcgtg gaaacagcca catcatagat
     5761 gattgcaacc tgcttacggg ggatgccctt ctccagcaat cgccgcattt gctgccatgt
     5821 ttcttcttgg tatttaggcc gacgcccacc tatacgacct tctgcgcgag ctgcatcaag
     5881 tccagcgcgt gtacgttcaa cgataagctc acgttccatt tctgccagcg cccccattac
     5941 gtgaaagaaa aagcgcccca ttggtgtact ggtgtcgatg gagtcagtga gactccggaa
     6001 gttaatgcct ctgtcacgca gctcttccac cagcacaact aagtgacgca tgctgcgccc
     6061 aagacggtct aacttccata cgaccagggt atcacctctg gaaagttacg gagtaccttt
     6121 tttaacccag ggcgctcagc ctttttgccg ctcgccttgt cctcaaaaat tagctcacat
     6181 cctgcgcttt caagagcgtt tcgttgtaaa gcagtgtttt gttcatttgt tgatacgcgt
     6241 acatagccta ttagagatcc tataaacgca gaaaggccca cccgaaggtg agccagtgtg
     6301 actctagtag agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga
     6361 ctgagccttt cgttttattt gatgcctgga gatccttatt gtgagtctgg ctggttagtg
     6421 gccatgagat attcgattgt gtcagtgaga tcatccaggt cgtcttgggt gatgcggtac
     6481 tcctgattgg atatctttga gtagtgttca gcaatggcgc gggcagcgtc ggtttcggcg
     6541 gggtctacag ataaagcgtt agagcaatgt ctaacgtcgt cgatggttgg tggaatgaaa
     6601 gccataatta tgcctcactg taagatccgg gtatggagaa acagtagaga gttgcgataa
     6661 aaagcgtcag gtaagatccg ctaatcttat ggataaaaat gctatggcat agcaaagtgt
     6721 gacgccgtgc aaataatcaa tgtggacttt tctgccgtga ttatagacac ttttgttacg
     6781 cgtttttgtc atggctttgg tcccgctttg ttacagaatg cttttaataa gcggggttac
     6841 cggtttggtt agcgagaaga gccagtaaaa gacgcagtga cggcaatgtc tgatgcaata
     6901 tggacaattg gtttcttctc tgaatggcgg gagtatgaaa agtatggctg aagcgcaaaa
     6961 tgatcccctg ctgccgggat actcgtttaa tgcccatctg gtggcgggtt taacgccgat
     7021 tgaggccaac ggttatctcg atttttttat cgaccgaccg ctgggaatga aaggttatat
     7081 tctcaatctc accattcgcg gtcagggggt ggtgaaaaat cagggacgag aatttgtttg
     7141 ccgaccgggt gatattttgc tgttcccgcc aggagagatt catcactacg gtcgtcatcc
     7201 ggaggctcgc gaatggtatc accagtgggt ttactttcgt ccgcgcgcct actggcatga
     7261 atggcttaac tggccgtcaa tatttgccaa tacggggttc tttcgcccgg atgaagcgca
     7321 ccagccgcat ttcagcgacc tgtttgggca aatcattaac gccgggcaag gggaagggcg
     7381 ctattcggag ctgctggcga taaatctgct tgagcaattg ttgctgcggc gcatggaagc
     7441 gattaacgag tcgctccatc caccgatgga taatcgggta cgcgaggctt gtcagtacat
     7501 cagcgatcac ctggcagaca gcaattttga tatcgccagc gtcgcacagc atgtttgctt
     7561 gtcgccgtcg cgtctgtcac gtcttttccg ccagcagtta gggattagcg tcttaagctg
     7621 gcgcgaggac caacgtatca gccaggcgaa gctgcttttg agcaccaccc ggatgcctat
     7681 cgccaccgtc ggtcgcaatg ttggttttga cgatcaactc tatttctcgc gggtatttaa
     7741 aaaatgcacc ggggccagcc cgagcgagtt ccgtgccggt tgtgaagaaa aagtgaatga
     7801 tgtagccgtc aagttgtcat aattggtaac gaatcagaca attgacggct cgatggagta
     7861 gcatagagat ctcatgaatt ccagaaatca tccttagcga aagctaagga ttttttttat
     7921 ctgaaattct gcctcgtgat accaattcgg agcctgcttt tttgtacaaa cttgttgata
     7981 atggcaattc ttgatcgggc acgtaagagg ttccaacttt caccataatg aaataagatc
     8041 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     8101 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     8161 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     8221 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     8281 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     8341 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     8401 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     8461 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     8521 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     8581 tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa
     8641 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg
     8701 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaatttga
     8761 tatcgagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     8821 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcctcg
     8881 agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg
     8941 agagtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca
     9001 gaatatgtga tacaggatat attccgcttc ctcgctcact ggctcgctac gctcggtcgt
     9061 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     9121 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     9181 ccccctgaca agcatcacga aatctgacgc tcagtctgaa tgacctgtca cgggataatc
     9241 cgaagtggtc agactggaaa atcagagggc aggaactgct gaacagcaaa aagtcagata
     9301 gcaccacata gcagacccgc cataaaacgc cctgagaagc ccgtgacggg cttttcttgt
     9361 attatgggta gtttccttgc atgaatccat aaaaggcgcc tgtagtgcca tttaccccca
     9421 ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg
     9481 tgcctgatgg tcggagacaa aaggaatatt cagcgatttg cccgagcttg cgagggtgct
     9541 acttaagcct ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat
     9601 ccttttgtaa tactgcggaa ctgactaaag tagt
//
