LOCUS       sequence-212802-        5368 bp    DNA     circular SYN 15-APR-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 Apr 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_N99K
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(228..322)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(353..1147)
                     /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(464..483)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1074..1093
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1290..2168)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2195..2479
                     /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     2402..2421
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2529..3206
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3239..3310
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3326..3353
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3884..4834)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTKYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 tacccataat acaagaaaag cccgtcacgg gcttctcagg gcgttttatg gcgggtctgc
       61 tatgtggtgc tatctgactt tttgctgttc agcagttcct gccctctgat tttccagtct
      121 gaccacttcg gattatcccg tgacaggtca ttcagactgg ctaatgcacc cagtaaggca
      181 gcggtatcat caacaggctt acccgtctta ctgtccctag tgcttggatt ctcaccaata
      241 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
      301 actggatcta tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga
      361 actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa
      421 gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca
      481 acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa
      541 agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat
      601 cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct
      661 gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc
      721 gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca
      781 gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca
      841 ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa
      901 cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct
      961 cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc
     1021 cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt
     1081 catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt
     1141 caatcatgcg aaacgatcct catcctgtct cttgatcaga tcatgatccc ctgcgccatc
     1201 agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca accttaccag
     1261 agggcgcccc agctggcaat tccgacgtct tatgacaact tgacggctac atcattcact
     1321 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     1381 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     1441 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     1501 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     1561 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     1621 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     1681 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     1741 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     1801 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     1861 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     1921 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     1981 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     2041 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     2101 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     2161 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
     2221 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
     2281 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
     2341 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
     2401 tatgccatag catttttatc cataagatta gcggattcta cctgacgctt tttatcgcaa
     2461 ctctctactg tttctccata cccgtttttt tgggaattca aaagatcttt taagaaggag
     2521 atatacatat ggcgagtagc gaagacgtta tcaaagagtt catgcgtttc aaagttcgta
     2581 tggaaggttc cgttaacggt cacgagttcg aaatcgaagg tgaaggtgaa ggtcgtccgt
     2641 acgaaggtac ccagaccgct aaactgaaag ttaccaaagg tggtccgctg ccgttcgctt
     2701 gggacatcct gtccccgcag ttccagtacg gttccaaagc ttacgttaaa cacccggctg
     2761 acatcccgga ctacctgaaa ctgtccttcc cggaaggttt caaatgggaa cgtgttatga
     2821 acttcgaaga cggtggtgtt gttaccgtta cccaggactc ctccctgcaa gacggtgagt
     2881 tcatctacaa agttaaactg cgtggtacca acttcccgtc cgacggtccg gttatgcaga
     2941 aaaaaaccat gggttgggaa gcttccaccg aacgtatgta cccggaagac ggtgctctga
     3001 aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg tcactacgac gctgaagtta
     3061 aaaccaccta catggctaaa aaaccggttc agctgccggg tgcttacaaa accgacatca
     3121 aactggacat cacctcccac aacgaagact acaccatcgt tgaacagtac gaacgtgctg
     3181 aaggtcgtca ctccaccggt gcttaaggat ccaaactcga gtaaggatct ccaggcatca
     3241 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3301 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac
     3361 ctagggtacg ggttttgctg cccgcaaacg ggctgttctg gtgttgctag tttgttatca
     3421 gaatcgcaga tccggcttca gccggtttgc cggctgaaag cgctatttct tccagaattg
     3481 ccatgatttt ttccccacgg gaggcgtcac tggctcccgt gttgtcggca gctttgattc
     3541 gataagcagc atcgcctgtt tcaggctgtc tatgtgtgac tgttgagctg taacaagttg
     3601 tctcaggtgt tcaatttcat gttctagttg ctttgtttta ctggtttcac ctgttctatt
     3661 aggtgttaca tgctgttcat ctgttacatt gtcgatctgt tcatggtgaa cagctttgaa
     3721 tgcaccaaaa actcgtaaaa gctctgatgt atctatcttt tttacaccgt tttcatctgt
     3781 gcatatggac agttttccct ttgatatgta acggtgaaca gttgttctac ttttgtttgt
     3841 tagtcttgat gcttcactga tagatacaag agccataaga acctcagatc cttccgtatt
     3901 tagccagtat gttctctagt gtggttcgtt gtttttgcgt gagccatgag aacgaaccat
     3961 tgagatcata cttactttgc atgtcactca aaaattttgc ctcaaaactg gtgagctgaa
     4021 tttttgcagt taaagcatcg tgtagtgttt ttcttagtcc gttatgtagg taggaatctg
     4081 atgtaatggt tgttggtatt ttgtcaccat tcatttttat ctggttgttc tcaagttcgg
     4141 ttacgagatc catttgtcta tctagttcaa cttggaaaat caacgtatca gtcgggcggc
     4201 ctcgcttatc aaccaccaat ttcatattgc tgtaagtgtt taaatcttta cttattggtt
     4261 tcaaaaccca ttggttaagc cttttaaact catggtagtt attttcaagc attaacatga
     4321 acttaaattc atcaaggcta atctctatat ttgccttgtg agttttcttt tgtgttagtt
     4381 cttttaataa ccactcataa atcctcatag agtatttgtt ttcaaaagac ttaacatgtt
     4441 ccagattata ttttatgaat ttttttaact ggaaaagata aggcaatatc tcttcactaa
     4501 aaactaattc taatttttcg cttgagaact tggcatactt tgtccactgg aaaatctcaa
     4561 agcctttaac caaaggattc ctgatttcca cagttctcgt catcagctct ctggttgctt
     4621 tagctaatac accataagca ttttccctac tgatgttcat catctgagcg tattggttat
     4681 aagtgaacga taccgtccgt tctttccttg tagggttttc aatcgtgggg ttgagtagtg
     4741 ccacacagca taaaattagc ttggtttcat gctccgttaa gtcatagcga ctaatcgcta
     4801 gttcatttgc tttgaaaaca actaattcag acatacatct caattggtct aggtgatttt
     4861 aatcactata ccaattgaga tgggctagtc aatgataatt actagtcctt ttcccgggtg
     4921 atctgggtat ctgtaaattc tgctagacct ttgctggaaa acttgtaaat tctgctagac
     4981 cctctgtaaa ttccgctaga cctttgtgtg ttttttttgt ttatattcaa gtggttataa
     5041 tttatagaat aaagaaagaa taaaaaaaga taaaaagaat agatcccagc cctgtgtata
     5101 actcactact ttagtcagtt ccgcagtatt acaaaaggat gtcgcaaacg ctgtttgctc
     5161 ctctacaaaa cagaccttaa aaccctaaag gcttaagtag caccctcgca agctcgggca
     5221 aatcgctgaa tattcctttt gtctccgacc atcaggcacc tgagtcgctg tctttttcgt
     5281 gacattcagt tcgctgcgct cacggctctg gcagtgaatg ggggtaaatg gcactacagg
     5341 cgccttttat ggattcatgc aaggaaac
//
