LOCUS       sequence-257185-        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"
     CDS             complement(7..885)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        912..1196
                     /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     1119..1138
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1246..1923
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1956..2027
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2043..2070
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2601..3551)
                     /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(4313..4407)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4438..5232)
                     /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(4549..4568)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5159..5178
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacgtcttat gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa
       61 ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc
      121 aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt
      181 cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg
      241 gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc
      301 aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc
      361 catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag
      421 cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg
      481 cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt
      541 ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac
      601 ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg
      661 cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac
      721 cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat
      781 aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt
      841 aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc
      901 gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt
      961 ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca
     1021 aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa
     1081 agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat
     1141 aagattagcg gattctacct gacgcttttt atcgcaactc tctactgttt ctccataccc
     1201 gtttttttgg gaattcaaaa gatcttttaa gaaggagata tacatatggc gagtagcgaa
     1261 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
     1321 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
     1381 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
     1441 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
     1501 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
     1561 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
     1621 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
     1681 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
     1741 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
     1801 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
     1861 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
     1921 taaggatcca aactcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
     1981 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2041 ctggctcacc ttcgggtggg cctttctgcg tttataccta gggtacgggt tttgctgccc
     2101 gcaaacgggc tgttctggtg ttgctagttt gttatcagaa tcgcagatcc ggcttcagcc
     2161 ggtttgccgg ctgaaagcgc tatttcttcc agaattgcca tgattttttc cccacgggag
     2221 gcgtcactgg ctcccgtgtt gtcggcagct ttgattcgat aagcagcatc gcctgtttca
     2281 ggctgtctat gtgtgactgt tgagctgtaa caagttgtct caggtgttca atttcatgtt
     2341 ctagttgctt tgttttactg gtttcacctg ttctattagg tgttacatgc tgttcatctg
     2401 ttacattgtc gatctgttca tggtgaacag ctttgaatgc accaaaaact cgtaaaagct
     2461 ctgatgtatc tatctttttt acaccgtttt catctgtgca tatggacagt tttccctttg
     2521 atatgtaacg gtgaacagtt gttctacttt tgtttgttag tcttgatgct tcactgatag
     2581 atacaagagc cataagaacc tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg
     2641 gttcgttgtt tttgcgtgag ccatgagaac gaaccattga gatcatactt actttgcatg
     2701 tcactcaaaa attttgcctc aaaactggtg agctgaattt ttgcagttaa agcatcgtgt
     2761 agtgtttttc ttagtccgtt atgtaggtag gaatctgatg taatggttgt tggtattttg
     2821 tcaccattca tttttatctg gttgttctca agttcggtta cgagatccat ttgtctatct
     2881 agttcaactt ggaaaatcaa cgtatcagtc gggcggcctc gcttatcaac caccaatttc
     2941 atattgctgt aagtgtttaa atctttactt attggtttca aaacccattg gttaagcctt
     3001 ttaaactcat ggtagttatt ttcaagcatt aacatgaact taaattcatc aaggctaatc
     3061 tctatatttg ccttgtgagt tttcttttgt gttagttctt ttaataacca ctcataaatc
     3121 ctcatagagt atttgttttc aaaagactta acatgttcca gattatattt tatgaatttt
     3181 tttaactgga aaagataagg caatatcttt tcactaaaaa ctaattctaa tttttcgctt
     3241 gagaactcgg catagtttgt ccactggaaa atctcaaagc ctttaaccaa aggattcctg
     3301 atttccacag ttctcgtcat cagctctctg gttgctttag ctaatacacc ataagcattt
     3361 tccctactga tgttcatcat ctgagcgtat tggttataag tgaacgatac cgtccgttct
     3421 ttccttgtag ggttttcaat cgtggggttg agtagtgcca cacagcataa aattagcttg
     3481 gtttcatgct ccgttaagtc atagcgacta atcgctagtt catttgcttt gaaaacaact
     3541 aattcagaca tacatctcaa ttggtctagg tgattttaat cactatacca attgagatgg
     3601 gctagtcaat gataattact agtccttttc ccgggtgatc tgggtatctg taaattctgc
     3661 tagacctttg ctggaaaact tgtaaattct gctagaccct ctgtaaattc cgctagacct
     3721 ttgtgtgttt tttttgttta tattcaagtg gttataattt atagaataaa gaaagaataa
     3781 aaaaagataa aaagaataga tcccagccct gtgtataact cactacttta gtcagttccg
     3841 cagtattaca aaaggatgtc gcaaacgctg tttgctcctc tacaaaacag accttaaaac
     3901 cctaaaggct taagtagcac cctcgcaagc tcgggcaaat cgctgaatat tccttttgtc
     3961 tccgaccatc aggcacctga gtcgctgtct ttttcgtgac attcagttcg ctgcgctcac
     4021 ggctctggca gtgaatgggg gtaaatggca ctacaggcgc cttttatgga ttcatgcaag
     4081 gaaactaccc ataatacaag aaaagcccgt cacgggcttc tcagggcgtt ttatggcggg
     4141 tctgctatgt ggtgctatct gactttttgc tgttcagcag ttcctgccct ctgattttcc
     4201 agtctgacca cttcggatta tcccgtgaca ggtcattcag actggctaat gcacccagta
     4261 aggcagcggt atcatcaaca ggcttacccg tcttactgtc cctagtgctt ggattctcac
     4321 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     4381 tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgctca
     4441 gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc
     4501 gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt
     4561 agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc
     4621 agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac
     4681 gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag
     4741 cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg
     4801 tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt
     4861 atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga
     4921 tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt
     4981 gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc
     5041 tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg
     5101 gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc
     5161 ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc
     5221 ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcatg atcccctgcg
     5281 ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt
     5341 accagagggc gccccagctg gcaattcc
//
