LOCUS       sequence-212796-        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_R46W
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      64..158
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             920..1870
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     WTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2401..2428)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2444..2515)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2548..3225)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3275..3559)
                     /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(3333..3352)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3586..4464
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4607..5368,1..33)
                     /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(4661..4680)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5271..5290
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gagagctcgc
       61 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      121 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg gacagtaaga
      181 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
      241 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
      301 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
      361 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
      421 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
      481 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
      541 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
      601 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
      661 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
      721 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
      781 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca gatcacccgg
      841 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
      901 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
      961 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     1021 caaccccacg attgaaaacc ctacaaggaa agaatggacg gtatcgttca cttataacca
     1081 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     1141 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
     1201 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     1261 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     1321 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     1381 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     1441 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     1501 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     1561 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     1621 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     1681 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     1741 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     1801 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     1861 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     1921 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     1981 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     2041 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     2101 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     2161 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     2221 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     2281 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     2341 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccc taggtataaa
     2401 cgcagaaagg cccacccgaa ggtgagccag tgtgactcta gtagagagcg ttcaccgaca
     2461 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     2521 tggagatcct tactcgagtt tggatcctta agcaccggtg gagtgacgac cttcagcacg
     2581 ttcgtactgt tcaacgatgg tgtagtcttc gttgtgggag gtgatgtcca gtttgatgtc
     2641 ggttttgtaa gcacccggca gctgaaccgg ttttttagcc atgtaggtgg ttttaacttc
     2701 agcgtcgtag tgaccaccgt ctttcagttt cagacgcatt ttgatttcac ctttcagagc
     2761 accgtcttcc gggtacatac gttcggtgga agcttcccaa cccatggttt ttttctgcat
     2821 aaccggaccg tcggacggga agttggtacc acgcagttta actttgtaga tgaactcacc
     2881 gtcttgcagg gaggagtcct gggtaacggt aacaacacca ccgtcttcga agttcataac
     2941 acgttcccat ttgaaacctt ccgggaagga cagtttcagg tagtccggga tgtcagccgg
     3001 gtgtttaacg taagctttgg aaccgtactg gaactgcggg gacaggatgt cccaagcgaa
     3061 cggcagcgga ccacctttgg taactttcag tttagcggtc tgggtacctt cgtacggacg
     3121 accttcacct tcaccttcga tttcgaactc gtgaccgtta acggaacctt ccatacgaac
     3181 tttgaaacgc atgaactctt tgataacgtc ttcgctactc gccatatgta tatctccttc
     3241 ttaaaagatc ttttgaattc ccaaaaaaac gggtatggag aaacagtaga gagttgcgat
     3301 aaaaagcgtc aggtagaatc cgctaatctt atggataaaa atgctatggc atagcaaagt
     3361 gtgacgccgt gcaaataatc aatgtggact tttctgccgt gattatagac acttttgtta
     3421 cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa tgcttttaat aagcggggtt
     3481 accggtttgg ttagcgagaa gagccagtaa aagacgcagt gacggcaatg tctgatgcaa
     3541 tatggacaat tggtttcttc tctgaatggc gggagtatga aaagtatggc tgaagcgcaa
     3601 aatgatcccc tgctgccggg atactcgttt aatgcccatc tggtggcggg tttaacgccg
     3661 attgaggcca acggttatct cgattttttt atcgaccgac cgctgggaat gaaaggttat
     3721 attctcaatc tcaccattcg cggtcagggg gtggtgaaaa atcagggacg agaatttgtt
     3781 tgccgaccgg gtgatatttt gctgttcccg ccaggagaga ttcatcacta cggtcgtcat
     3841 ccggaggctc gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc ctactggcat
     3901 gaatggctta actggccgtc aatatttgcc aatacggggt tctttcgccc ggatgaagcg
     3961 caccagccgc atttcagcga cctgtttggg caaatcatta acgccgggca aggggaaggg
     4021 cgctattcgg agctgctggc gataaatctg cttgagcaat tgttactgcg gcgcatggaa
     4081 gcgattaacg agtcgctcca tccaccgatg gataatcggg tacgcgaggc ttgtcagtac
     4141 atcagcgatc acctggcaga cagcaatttt gatatcgcca gcgtcgcaca gcatgtttgc
     4201 ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt tagggattag cgtcttaagc
     4261 tggcgcgagg accaacgtat cagccaggcg aagctgcttt tgagcaccac ccggatgcct
     4321 atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac tctatttctc gcgggtattt
     4381 aaaaaatgca ccggggccag cccgagcgag ttccgtgccg gttgtgaaga aaaagtgaat
     4441 gatgtagccg tcaagttgtc ataagacgtc ggaattgcca gctggggcgc cctctggtaa
     4501 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     4561 caggggatca tgatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4621 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4681 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4741 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4801 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4861 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4921 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4981 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     5041 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     5101 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     5161 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     5221 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     5281 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     5341 tatcgccgct cccgattcgc agcgcatc
//
