LOCUS       sequence-212801-        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_K102E
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1068..1162)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1193..1987)
                     /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(1304..1323)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1914..1933
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2130..3008)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3035..3319
                     /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     3242..3261
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3369..4046
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4079..4150
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4166..4193
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(join(4724..5368,1..306))
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAEFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 ctcggcatag tttgtccact ggaaaatctc aaagccttta accaaaggat tcctgatttc
       61 cacagttctc gtcatcagct ctctggttgc tttagctaat acaccataag cattttccct
      121 actgatgttc atcatctgag cgtattggtt ataagtgaac gataccgtcc gttctttcct
      181 tgtagggttt tcaatcgtgg ggttgagtag tgccacacag cataaaatta gcttggtttc
      241 atgctccgtt aagtcatagc gactaatcgc tagttcattt gctttgaaaa caactaattc
      301 agacatacat ctcaattggt ctaggtgatt ttaatcacta taccaattga gatgggctag
      361 tcaatgataa ttactagtcc ttttcccggg tgatctgggt atctgtaaat tctgctagac
      421 ctttgctgga aaacttgtaa attctgctag accctctgta aattccgcta gacctttgtg
      481 tgtttttttt gtttatattc aagtggttat aatttataga ataaagaaag aataaaaaaa
      541 gataaaaaga atagatccca gccctgtgta taactcacta ctttagtcag ttccgcagta
      601 ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa aacagacctt aaaaccctaa
      661 aggcttaagt agcaccctcg caagctcggg caaatcgctg aatattcctt ttgtctccga
      721 ccatcaggca cctgagtcgc tgtctttttc gtgacattca gttcgctgcg ctcacggctc
      781 tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt atggattcat gcaaggaaac
      841 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
      901 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
      961 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
     1021 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
     1081 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     1141 actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga
     1201 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
     1261 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
     1321 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
     1381 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
     1441 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
     1501 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
     1561 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
     1621 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
     1681 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
     1741 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
     1801 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     1861 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     1921 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     1981 caatcatgcg aaacgatcct catcctgtct cttgatcaga tcatgatccc ctgcgccatc
     2041 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     2101 agggcgcccc agctggcaat tccgacgtct tatgacaact tgacggctac atcattcact
     2161 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     2221 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     2281 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     2341 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     2401 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     2461 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     2521 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     2581 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     2641 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     2701 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     2761 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     2821 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     2881 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     2941 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     3001 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
     3061 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
     3121 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
     3181 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
     3241 tatgccatag catttttatc cataagatta gcggattcta cctgacgctt tttatcgcaa
     3301 ctctctactg tttctccata cccgtttttt tgggaattca aaagatcttt taagaaggag
     3361 atatacatat ggcgagtagc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
     3421 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
     3481 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
     3541 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
     3601 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
     3661 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
     3721 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
     3781 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
     3841 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
     3901 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
     3961 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
     4021 aaggtcgtca ctccaccggt gcttaaggat ccaaactcga gtaaggatct ccaggcatca
     4081 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     4141 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac
     4201 ctagggtacg ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca
     4261 gaatcgcaga tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg
     4321 ccatgatttt ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc
     4381 gataagcagc atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg
     4441 tctcaggtgt tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt
     4501 aggtgttaca tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa
     4561 tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt
     4621 gcatatggac agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt
     4681 tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt
     4741 tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat
     4801 tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa
     4861 tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg
     4921 atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg
     4981 ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc
     5041 ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt
     5101 tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga
     5161 acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt
     5221 cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt
     5281 ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa
     5341 aaactaattc taatttttcg cttgagaa
//
