LOCUS       sequence-212806-        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_E115K
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      95..189
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             951..1901
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAEFS
                     SEKLELVFSEKILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2432..2459)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2475..2546)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2579..3256)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3306..3590)
                     /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(3364..3383)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3617..4495
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4638..5368,1..64)
                     /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(4692..4711)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5302..5321
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
       61 ctgagcggga ctctggggtt cgagagctcg cttggactcc tgttgataga tccagtaatg
      121 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      181 ggtgagaatc caagcactag ggacagtaag acgggtaagc ctgttgatga taccgctgcc
      241 ttactgggtg cattagccag tctgaatgac ctgtcacggg ataatccgaa gtggtcagac
      301 tggaaaatca gagggcagga actgctgaac agcaaaaagt cagatagcac cacatagcag
      361 acccgccata aaacgccctg agaagcccgt gacgggcttt tcttgtatta tgggtagttt
      421 ccttgcatga atccataaaa ggcgcctgta gtgccattta cccccattca ctgccagagc
      481 cgtgagcgca gcgaactgaa tgtcacgaaa aagacagcga ctcaggtgcc tgatggtcgg
      541 agacaaaagg aatattcagc gatttgcccg agcttgcgag ggtgctactt aagcctttag
      601 ggttttaagg tctgttttgt agaggagcaa acagcgtttg cgacatcctt ttgtaatact
      661 gcggaactga ctaaagtagt gagttataca cagggctggg atctattctt tttatctttt
      721 tttattcttt ctttattcta taaattataa ccacttgaat ataaacaaaa aaaacacaca
      781 aaggtctagc ggaatttaca gagggtctag cagaatttac aagttttcca gcaaaggtct
      841 agcagaattt acagataccc agatcacccg ggaaaaggac tagtaattat cattgactag
      901 cccatctcaa ttggtatagt gattaaaatc acctagacca attgagatgt atgtctgaat
      961 tagttgtttt caaagcaaat gaactagcga ttagtcgcta tgacttaacg gagcatgaaa
     1021 ccaagctaat tttatgctgt gtggcactac tcaaccccac gattgaaaac cctacaagga
     1081 aagaacggac ggtatcgttc acttataacc aatacgctca gatgatgaac atcagtaggg
     1141 aaaatgctta tggtgtatta gctaaagcaa ccagagagct gatgacgaga actgtggaaa
     1201 tcaggaatcc tttggttaaa ggctttgaga ttttccagtg gacaaactat gccgagttct
     1261 caagcgaaaa attagaatta gtttttagtg aaaagatatt gccttatctt ttccagttaa
     1321 aaaaattcat aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa tactctatga
     1381 ggatttatga gtggttatta aaagaactaa cacaaaagaa aactcacaag gcaaatatag
     1441 agattagcct tgatgaattt aagttcatgt taatgcttga aaataactac catgagttta
     1501 aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact tacagcaata
     1561 tgaaattggt ggttgataag cgaggccgcc cgactgatac gttgattttc caagttgaac
     1621 tagatagaca aatggatctc gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg
     1681 acaaaatacc aacaaccatt acatcagatt cctacctaca taacggacta agaaaaacac
     1741 tacacgatgc tttaactgca aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg
     1801 acatgcaaag taagtatgat ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa
     1861 ccacactaga gaacatactg gctaaatacg gaaggatctg aggttcttat ggctcttgta
     1921 tctatcagtg aagcatcaag actaacaaac aaaagtagaa caactgttca ccgttacata
     1981 tcaaagggaa aactgtccat atgcacagat gaaaacggtg taaaaaagat agatacatca
     2041 gagcttttac gagtttttgg tgcattcaaa gctgttcacc atgaacagat cgacaatgta
     2101 acagatgaac agcatgtaac acctaataga acaggtgaaa ccagtaaaac aaagcaacta
     2161 gaacatgaaa ttgaacacct gagacaactt gttacagctc aacagtcaca catagacagc
     2221 ctgaaacagg cgatgctgct tatcgaatca aagctgccga caacacggga gccagtgacg
     2281 cctcccgtgg ggaaaaaatc atggcaattc tggaagaaat agcgctttca gccggcaaac
     2341 cggctgaagc cggatctgcg attctgataa caaactagca acaccagaac agcccgtttg
     2401 cgggcagcaa aacccgtacc ctaggtataa acgcagaaag gcccacccga aggtgagcca
     2461 gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2521 ttcgactgag cctttcgttt tatttgatgc ctggagatcc ttactcgagt ttggatcctt
     2581 aagcaccggt ggagtgacga ccttcagcac gttcgtactg ttcaacgatg gtgtagtctt
     2641 cgttgtggga ggtgatgtcc agtttgatgt cggttttgta agcacccggc agctgaaccg
     2701 gttttttagc catgtaggtg gttttaactt cagcgtcgta gtgaccaccg tctttcagtt
     2761 tcagacgcat tttgatttca cctttcagag caccgtcttc cgggtacata cgttcggtgg
     2821 aagcttccca acccatggtt tttttctgca taaccggacc gtcggacggg aagttggtac
     2881 cacgcagttt aactttgtag atgaactcac cgtcttgcag ggaggagtcc tgggtaacgg
     2941 taacaacacc accgtcttcg aagttcataa cacgttccca tttgaaacct tccgggaagg
     3001 acagtttcag gtagtccggg atgtcagccg ggtgtttaac gtaagctttg gaaccgtact
     3061 ggaactgcgg ggacaggatg tcccaagcga acggcagcgg accacctttg gtaactttca
     3121 gtttagcggt ctgggtacct tcgtacggac gaccttcacc ttcaccttcg atttcgaact
     3181 cgtgaccgtt aacggaacct tccatacgaa ctttgaaacg catgaactct ttgataacgt
     3241 cttcgctact cgccatatgt atatctcctt cttaaaagat cttttgaatt cccaaaaaaa
     3301 cgggtatgga gaaacagtag agagttgcga taaaaagcgt caggtagaat ccgctaatct
     3361 tatggataaa aatgctatgg catagcaaag tgtgacgccg tgcaaataat caatgtggac
     3421 ttttctgccg tgattataga cacttttgtt acgcgttttt gtcatggctt tggtcccgct
     3481 ttgttacaga atgcttttaa taagcggggt taccggtttg gttagcgaga agagccagta
     3541 aaagacgcag tgacggcaat gtctgatgca atatggacaa ttggtttctt ctctgaatgg
     3601 cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg gatactcgtt
     3661 taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc tcgatttttt
     3721 tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc gcggtcaggg
     3781 ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt tgctgttccc
     3841 gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt atcaccagtg
     3901 ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt caatatttgc
     3961 caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg acctgtttgg
     4021 gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg cgataaatct
     4081 gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc atccaccgat
     4141 ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag acagcaattt
     4201 tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt cacatctttt
     4261 ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta tcagccaggc
     4321 gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca atgttggttt
     4381 tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca gcccgagcga
     4441 gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt cataagacgt
     4501 cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga
     4561 tggctttctt gccgccaagg atctgatggc gcaggggatc atgatctgat caagagacag
     4621 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4681 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4741 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4801 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4861 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4921 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4981 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     5041 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     5101 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     5161 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     5221 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     5281 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     5341 aatgggctga ccgcttcctc gtgcttta
//
