LOCUS       sequence-212793-        5368 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  RepA Mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5368)
  AUTHORS   Thompson MG, Sedaghatian N, Barajas JF, Wehrs M, Bailey CB, Kaplan 
            N, Hillson NJ, Mukhopadhyay A, Keasling JD
  TITLE     Isolation and characterization of novel mutations in the pSC101 
            origin that increase copy number.
  JOURNAL   Sci Rep. 2018 Jan 25;8(1):1590. doi: 10.1038/s41598-018-20016-w.
  PUBMED    29371642
REFERENCE   2  (bases 1 to 5368)
  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 Rep."; 
            date: "2018-01-25"; volume: "8(1):1590. doi"; pages: " 
            10.1038/s41598-018-20016-w"
COMMENT     Sequence Label: pBbS8k-RFP_M78I
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..49
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      191..285
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1047..1997
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELITRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2528..2555)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2571..2642)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2675..3352)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3402..3686)
                     /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(3460..3479)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3713..4591
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4734..5368,1..160)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4788..4807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
       61 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
      121 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgag
      181 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
      241 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag cactagggac
      301 agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg
      361 aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg
      421 ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa
      481 gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg
      541 cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc
      601 acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt
      661 tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag
      721 gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt
      781 tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt attctataaa
      841 ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg
      901 gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag atacccagat
      961 cacccgggaa aaggactagt aattatcatt gactagccca tctcaattgg tatagtgatt
     1021 aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac
     1081 tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg
     1141 cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt
     1201 ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta
     1261 aagcaaccag agagctgata acgagaactg tggaaatcag gaatcctttg gttaaaggct
     1321 ttgagatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt
     1381 ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg
     1441 aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag
     1501 aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt
     1561 tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga
     1621 aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag
     1681 gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa
     1741 ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat
     1801 cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta actgcaaaaa
     1861 ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag tatgatctca
     1921 atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta
     1981 aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta
     2041 acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact gtccatatgc
     2101 acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca
     2161 ttcaaagctg ttcaccatga acagatcgac aatgtaacag atgaacagca tgtaacacct
     2221 aatagaacag gtgaaaccag taaaacaaag caactagaac atgaaattga acacctgaga
     2281 caacttgtta cagctcaaca gtcacacata gacagcctga aacaggcgat gctgcttatc
     2341 gaatcaaagc tgccgacaac acgggagcca gtgacgcctc ccgtggggaa aaaatcatgg
     2401 caattctgga agaaatagcg ctttcagccg gcaaaccggc tgaagccgga tctgcgattc
     2461 tgataacaaa ctagcaacac cagaacagcc cgtttgcggg cagcaaaacc cgtaccctag
     2521 gtataaacgc agaaaggccc acccgaaggt gagccagtgt gactctagta gagagcgttc
     2581 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     2641 tgatgcctgg agatccttac tcgagtttgg atccttaagc accggtggag tgacgacctt
     2701 cagcacgttc gtactgttca acgatggtgt agtcttcgtt gtgggaggtg atgtccagtt
     2761 tgatgtcggt tttgtaagca cccggcagct gaaccggttt tttagccatg taggtggttt
     2821 taacttcagc gtcgtagtga ccaccgtctt tcagtttcag acgcattttg atttcacctt
     2881 tcagagcacc gtcttccggg tacatacgtt cggtggaagc ttcccaaccc atggtttttt
     2941 tctgcataac cggaccgtcg gacgggaagt tggtaccacg cagtttaact ttgtagatga
     3001 actcaccgtc ttgcagggag gagtcctggg taacggtaac aacaccaccg tcttcgaagt
     3061 tcataacacg ttcccatttg aaaccttccg ggaaggacag tttcaggtag tccgggatgt
     3121 cagccgggtg tttaacgtaa gctttggaac cgtactggaa ctgcggggac aggatgtccc
     3181 aagcgaacgg cagcggacca cctttggtaa ctttcagttt agcggtctgg gtaccttcgt
     3241 acggacgacc ttcaccttca ccttcgattt cgaactcgtg accgttaacg gaaccttcca
     3301 tacgaacttt gaaacgcatg aactctttga taacgtcttc gctactcgcc atatgtatat
     3361 ctccttctta aaagatcttt tgaattccca aaaaaacggg tatggagaaa cagtagagag
     3421 ttgcgataaa aagcgtcagg tagaatccgc taatcttatg gataaaaatg ctatggcata
     3481 gcaaagtgtg acgccgtgca aataatcaat gtggactttt ctgccgtgat tatagacact
     3541 tttgttacgc gtttttgtca tggctttggt cccgctttgt tacagaatgc ttttaataag
     3601 cggggttacc ggtttggtta gcgagaagag ccagtaaaag acgcagtgac ggcaatgtct
     3661 gatgcaatat ggacaattgg tttcttctct gaatggcggg agtatgaaaa gtatggctga
     3721 agcgcaaaat gatcccctgc tgccgggata ctcgtttaat gcccatctgg tggcgggttt
     3781 aacgccgatt gaggccaacg gttatctcga tttttttatc gaccgaccgc tgggaatgaa
     3841 aggttatatt ctcaatctca ccattcgcgg tcagggggtg gtgaaaaatc agggacgaga
     3901 atttgtttgc cgaccgggtg atattttgct gttcccgcca ggagagattc atcactacgg
     3961 tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt tactttcgtc cgcgcgccta
     4021 ctggcatgaa tggcttaact ggccgtcaat atttgccaat acggggttct ttcgcccgga
     4081 tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa atcattaacg ccgggcaagg
     4141 ggaagggcgc tattcggagc tgctggcgat aaatctgctt gagcaattgt tactgcggcg
     4201 catggaagcg attaacgagt cgctccatcc accgatggat aatcgggtac gcgaggcttg
     4261 tcagtacatc agcgatcacc tggcagacag caattttgat atcgccagcg tcgcacagca
     4321 tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc cagcagttag ggattagcgt
     4381 cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag ctgcttttga gcaccacccg
     4441 gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac gatcaactct atttctcgcg
     4501 ggtatttaaa aaatgcaccg gggccagccc gagcgagttc cgtgccggtt gtgaagaaaa
     4561 agtgaatgat gtagccgtca agttgtcata agacgtcgga attgccagct ggggcgccct
     4621 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
     4681 gatggcgcag gggatcatga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4741 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4801 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4861 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4921 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4981 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     5041 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     5101 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5161 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5221 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5281 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5341 tttctggatt catcgactgt ggccggct
//
