LOCUS       sequence-266113-        5295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Accessory plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Brodel AK, Rodrigues R, Jaramillo A, Isalan M
  TITLE     Accelerated evolution of a minimal 63-amino acid dual transcription 
            factor.
  JOURNAL   Sci Adv. 2020 Jun 10;6(24):eaba2728. doi: 10.1126/sciadv.aba2728. 
            eCollection 2020 Jun.
  PUBMED    32577520
REFERENCE   2  (bases 1 to 5295)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Adv."; 
            date: "2020-06-10"; volume: "6(24):eaba2728. doi"; pages: " 
            10.1126/sciadv.aba2728. eCollection 2020 Jun"
COMMENT     Sequence Label: pSC101-Delta-PS-PM,CS-RBSBBa_B0034-geneVI
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     471..493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     485..502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     486..502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     512..531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        512..530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     555..571
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     556..572
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             684..695
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             702..1040
                     /codon_start=1
                     /gene="VI"
                     /product="pVI"
                     /label=M13 gene VI
                     /note="minor coat protein"
                     /translation="MPVLLGIPLLLRFLGFLLVTLFGYLLTFLKKGFGKIAIAISLFLA
                     LIIGLNSILVGYLSDISAQLPSDFVQGVQLILPSNALPCFYVILSVKAAIFIFDVKQKI
                     VSYLDWDK"
     primer_bind     complement(1092..1112)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1092..1110)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1131..1147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1131..1147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1155..1171
                     /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)."
     promoter        complement(join(1179..1185,1186..1203,1204..1209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1362..1379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      2524..2746
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2794..3744
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(4782..5295,1..146))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
       61 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
      121 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
      181 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttaca
      241 aaaggatcta ggtgaagatc ctttttgata atctcatgag cggatacata tttgaatgta
      301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccaccttcag
      361 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
      421 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
      481 acgttgtaaa acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
      541 taccgggccc cccctcgagg tcgacggtat cgataagctt gatatcgaat tcgcaaccat
      601 tatcaccggc ggtgataaaa tagttatcac cggcggtgat agatattaca aactttcttg
      661 tatagattta caatgtatct tgtaaagagg agaaatacta gatgccagtt cttttgggta
      721 ttccgttatt attgcgtttc ctcggtttcc ttctggtaac tttgttcggc tatctgctta
      781 cttttcttaa aaagggcttc ggtaagatag ctattgctat ttcattgttt cttgctctta
      841 ttattgggct taactcaatt cttgtgggtt atctctctga tattagcgct caattaccct
      901 ctgactttgt tcagggtgtt cagttaattc tcccgtctaa tgcgcttccc tgtttttatg
      961 ttattctctc tgtaaaggct gctattttca tttttgacgt taaacaaaaa atcgtttctt
     1021 atttggattg ggataaataa cgcaaaaaac cccgcttcgg cggggttttt tcgcagctcc
     1081 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     1141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1441 tccacagaat caggggataa cgcaggaaag aacatgtgag atctctacgg gtcggatttg
     1501 aagtcgtctt ggtaggaggc agcctgaatg gcgaatgccg atgcccttga gagccttcaa
     1561 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     1621 cttctttatc atgcaactcg taggacaggg tgccggcagc gctctgggtc attttcggcg
     1681 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     1741 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     1801 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     1861 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     1921 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     1981 gatcgctcgc ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga
     2041 tttatgccgc ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat
     2101 accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa
     2161 tggaagccgg cggcacctcg ctaacggatt caccactccg cagacccgcc ataaaacgcc
     2221 ctgagaagcc cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata
     2281 aaaggcgcct gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact
     2341 gaatgtcacg aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc
     2401 agcgatttgc ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt
     2461 tgtagaggag caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt
     2521 agtgagttat acacagggct gggatctatt ctttttatct ttttttattc tttctttatt
     2581 ctataaatta taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt
     2641 acagagggtc tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata
     2701 cccacaactc aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat
     2761 agtgattaaa atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca
     2821 aatgaactag cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc
     2881 tgtgtggcac tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg
     2941 ttcacttata accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta
     3001 ttagctaaag caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt
     3061 aaaggctttg agatcttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa
     3121 ttagttttta gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat
     3181 aatctggaac atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta
     3241 ttaaaagaac taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa
     3301 tttaagttca tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg
     3361 gttttgaaac caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat
     3421 aagcgaggcc gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat
     3481 ctcgtaaccg aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc
     3541 attacatcag attcctacct acataacgga ctaagaaaaa cactacacga tgctttaaca
     3601 gcaaaaattc agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat
     3661 gatctcaatg gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata
     3721 ctggctaaat acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc
     3781 aagactaaca aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc
     3841 catatgcaca gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt
     3901 tggtgcattc aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt
     3961 aacacctaat agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca
     4021 cctgagacaa cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct
     4081 gcttatcgaa tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa
     4141 atcatggcaa ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct
     4201 gcgattctga taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt
     4261 acttttggac gttccggcgg ttttttgtgg cgagtggtgt tcgggcggtg cgcgcaagat
     4321 ccattatgtt aaacgggcga gtttacatct caaaaccgcc cgcttaacac catcagaaat
     4381 cctcagcgcg attttaagca ccaacccccc cccgtaacac ccaaatccat actgaaagtg
     4441 gctttgttga ataaatcaga tttcgggtaa gtctcccccg tagcgggttg tgttttcagg
     4501 caatacgcac gctttcaggc atacctgctt tcgtcatttt gttcagcgct cgtaccaggg
     4561 ccatagcctc cacaacctga ccatcgtagt cacgcagcgt cagtgaaccc ccgaacagag
     4621 atctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gacaataacc
     4681 ctgataaatg cttcaataat cgcagaccaa aacgatctca agaagatcat cttattaatc
     4741 agataaaata tttctaggca ccaataactg ccttaaaaaa attacgcccc gccctgccac
     4801 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     4861 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     4921 cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     4981 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     5041 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     5101 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     5161 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     5221 cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     5281 ttgtgcttat ttttc
//
