LOCUS       sequence-212803-        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_N99D
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      171..265
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1027..1977
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2508..2535)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2551..2622)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2655..3332)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3382..3666)
                     /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(3440..3459)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3693..4571
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4714..5368,1..140)
                     /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(4768..4787)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
       61 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
      121 ttcttgacga gttcttctga gcgggactct ggggttcgag agctcgcttg gactcctgtt
      181 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
      241 ccgggcgttt tttattggtg agaatccaag cactagggac agtaagacgg gtaagcctgt
      301 tgatgatacc gctgccttac tgggtgcatt agccagtctg aatgacctgt cacgggataa
      361 tccgaagtgg tcagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga
      421 tagcaccaca tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt
      481 gtattatggg tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc
      541 cattcactgc cagagccgtg agcgcagcga actgaatgtc acgaaaaaga cagcgactca
      601 ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg
      661 ctacttaagc ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac
      721 atccttttgt aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct
      781 attcttttta tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa
      841 acaaaaaaaa cacacaaagg tctagcggaa tttacagagg gtctagcaga atttacaagt
      901 tttccagcaa aggtctagca gaatttacag atacccagat cacccgggaa aaggactagt
      961 aattatcatt gactagccca tctcaattgg tatagtgatt aaaatcacct agaccaattg
     1021 agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac tagcgattag tcgctatgac
     1081 ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg cactactcaa ccccacgatt
     1141 gaaaacccta caaggaaaga acggacggta tcgttcactt ataaccaata cgctcagatg
     1201 atgaacatca gtagggaaaa tgcttatggt gtattagcta aagcaaccag agagctgatg
     1261 acgagaactg tggaaatcag gaatcctttg gttaaaggct ttgagatttt ccagtggaca
     1321 gactatgcca agttctcaag cgaaaaatta gaattagttt ttagtgaaga gatattgcct
     1381 tatcttttcc agttaaaaaa attcataaaa tataatctgg aacatgttaa gtcttttgaa
     1441 aacaaatact ctatgaggat ttatgagtgg ttattaaaag aactaacaca aaagaaaact
     1501 cacaaggcaa atatagagat tagccttgat gaatttaagt tcatgttaat gcttgaaaat
     1561 aactaccatg agtttaaaag gcttaaccaa tgggttttga aaccaataag taaagattta
     1621 aacacttaca gcaatatgaa attggtggtt gataagcgag gccgcccgac tgatacgttg
     1681 attttccaag ttgaactaga tagacaaatg gatctcgtaa ccgaacttga gaacaaccag
     1741 ataaaaatga atggtgacaa aataccaaca accattacat cagattccta cctacataac
     1801 ggactaagaa aaacactaca cgatgcttta actgcaaaaa ttcagctcac cagttttgag
     1861 gcaaaatttt tgagtgacat gcaaagtaag tatgatctca atggttcgtt ctcatggctc
     1921 acgcaaaaac aacgaaccac actagagaac atactggcta aatacggaag gatctgaggt
     1981 tcttatggct cttgtatcta tcagtgaagc atcaagacta acaaacaaaa gtagaacaac
     2041 tgttcaccgt tacatatcaa agggaaaact gtccatatgc acagatgaaa acggtgtaaa
     2101 aaagatagat acatcagagc ttttacgagt ttttggtgca ttcaaagctg ttcaccatga
     2161 acagatcgac aatgtaacag atgaacagca tgtaacacct aatagaacag gtgaaaccag
     2221 taaaacaaag caactagaac atgaaattga acacctgaga caacttgtta cagctcaaca
     2281 gtcacacata gacagcctga aacaggcgat gctgcttatc gaatcaaagc tgccgacaac
     2341 acgggagcca gtgacgcctc ccgtggggaa aaaatcatgg caattctgga agaaatagcg
     2401 ctttcagccg gcaaaccggc tgaagccgga tctgcgattc tgataacaaa ctagcaacac
     2461 cagaacagcc cgtttgcggg cagcaaaacc cgtaccctag gtataaacgc agaaaggccc
     2521 acccgaaggt gagccagtgt gactctagta gagagcgttc accgacaaac aacagataaa
     2581 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctgg agatccttac
     2641 tcgagtttgg atccttaagc accggtggag tgacgacctt cagcacgttc gtactgttca
     2701 acgatggtgt agtcttcgtt gtgggaggtg atgtccagtt tgatgtcggt tttgtaagca
     2761 cccggcagct gaaccggttt tttagccatg taggtggttt taacttcagc gtcgtagtga
     2821 ccaccgtctt tcagtttcag acgcattttg atttcacctt tcagagcacc gtcttccggg
     2881 tacatacgtt cggtggaagc ttcccaaccc atggtttttt tctgcataac cggaccgtcg
     2941 gacgggaagt tggtaccacg cagtttaact ttgtagatga actcaccgtc ttgcagggag
     3001 gagtcctggg taacggtaac aacaccaccg tcttcgaagt tcataacacg ttcccatttg
     3061 aaaccttccg ggaaggacag tttcaggtag tccgggatgt cagccgggtg tttaacgtaa
     3121 gctttggaac cgtactggaa ctgcggggac aggatgtccc aagcgaacgg cagcggacca
     3181 cctttggtaa ctttcagttt agcggtctgg gtaccttcgt acggacgacc ttcaccttca
     3241 ccttcgattt cgaactcgtg accgttaacg gaaccttcca tacgaacttt gaaacgcatg
     3301 aactctttga taacgtcttc gctactcgcc atatgtatat ctccttctta aaagatcttt
     3361 tgaattccca aaaaaacggg tatggagaaa cagtagagag ttgcgataaa aagcgtcagg
     3421 tagaatccgc taatcttatg gataaaaatg ctatggcata gcaaagtgtg acgccgtgca
     3481 aataatcaat gtggactttt ctgccgtgat tatagacact tttgttacgc gtttttgtca
     3541 tggctttggt cccgctttgt tacagaatgc ttttaataag cggggttacc ggtttggtta
     3601 gcgagaagag ccagtaaaag acgcagtgac ggcaatgtct gatgcaatat ggacaattgg
     3661 tttcttctct gaatggcggg agtatgaaaa gtatggctga agcgcaaaat gatcccctgc
     3721 tgccgggata ctcgtttaat gcccatctgg tggcgggttt aacgccgatt gaggccaacg
     3781 gttatctcga tttttttatc gaccgaccgc tgggaatgaa aggttatatt ctcaatctca
     3841 ccattcgcgg tcagggggtg gtgaaaaatc agggacgaga atttgtttgc cgaccgggtg
     3901 atattttgct gttcccgcca ggagagattc atcactacgg tcgtcatccg gaggctcgcg
     3961 aatggtatca ccagtgggtt tactttcgtc cgcgcgccta ctggcatgaa tggcttaact
     4021 ggccgtcaat atttgccaat acggggttct ttcgcccgga tgaagcgcac cagccgcatt
     4081 tcagcgacct gtttgggcaa atcattaacg ccgggcaagg ggaagggcgc tattcggagc
     4141 tgctggcgat aaatctgctt gagcaattgt tactgcggcg catggaagcg attaacgagt
     4201 cgctccatcc accgatggat aatcgggtac gcgaggcttg tcagtacatc agcgatcacc
     4261 tggcagacag caattttgat atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc
     4321 gtctgtcaca tcttttccgc cagcagttag ggattagcgt cttaagctgg cgcgaggacc
     4381 aacgtatcag ccaggcgaag ctgcttttga gcaccacccg gatgcctatc gccaccgtcg
     4441 gtcgcaatgt tggttttgac gatcaactct atttctcgcg ggtatttaaa aaatgcaccg
     4501 gggccagccc gagcgagttc cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca
     4561 agttgtcata agacgtcgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     4621 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcatga
     4681 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4741 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4801 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4861 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4921 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4981 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     5041 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     5101 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     5161 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5221 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5281 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5341 ggccggctgg gtgtggcgga ccgctatc
//
