LOCUS       sequence-212804-        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_E83K
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..28
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      170..264
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1026..1976
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVKIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2507..2534)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2550..2621)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2654..3331)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3381..3665)
                     /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(3439..3458)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3692..4570
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4713..5368,1..139)
                     /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(4767..4786)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
       61 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
      121 tcttgacgag ttcttctgag cgggactctg gggttcgaga gctcgcttgg actcctgttg
      181 atagatccag taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc
      241 cgggcgtttt ttattggtga gaatccaagc actagggaca gtaagacggg taagcctgtt
      301 gatgataccg ctgccttact gggtgcatta gccagtctga atgacctgtc acgggataat
      361 ccgaagtggt cagactggaa aatcagaggg caggaactgc tgaacagcaa aaagtcagat
      421 agcaccacat agcagacccg ccataaaacg ccctgagaag cccgtgacgg gcttttcttg
      481 tattatgggt agtttccttg catgaatcca taaaaggcgc ctgtagtgcc atttaccccc
      541 attcactgcc agagccgtga gcgcagcgaa ctgaatgtca cgaaaaagac agcgactcag
      601 gtgcctgatg gtcggagaca aaaggaatat tcagcgattt gcccgagctt gcgagggtgc
      661 tacttaagcc tttagggttt taaggtctgt tttgtagagg agcaaacagc gtttgcgaca
      721 tccttttgta atactgcgga actgactaaa gtagtgagtt atacacaggg ctgggatcta
      781 ttctttttat ctttttttat tctttcttta ttctataaat tataaccact tgaatataaa
      841 caaaaaaaac acacaaaggt ctagcggaat ttacagaggg tctagcagaa tttacaagtt
      901 ttccagcaaa ggtctagcag aatttacaga tacccagatc acccgggaaa aggactagta
      961 attatcattg actagcccat ctcaattggt atagtgatta aaatcaccta gaccaattga
     1021 gatgtatgtc tgaattagtt gttttcaaag caaatgaact agcgattagt cgctatgact
     1081 taacggagca tgaaaccaag ctaattttat gctgtgtggc actactcaac cccacgattg
     1141 aaaaccctac aaggaaagaa cggacggtat cgttcactta taaccaatac gctcagatga
     1201 tgaacatcag tagggaaaat gcttatggtg tattagctaa agcaaccaga gagctgatga
     1261 cgagaactgt gaaaatcagg aatcctttgg ttaaaggctt tgagattttc cagtggacaa
     1321 actatgccaa gttctcaagc gaaaaattag aattagtttt tagtgaagag atattgcctt
     1381 atcttttcca gttaaaaaaa ttcataaaat ataatctgga acatgttaag tcttttgaaa
     1441 acaaatactc tatgaggatt tatgagtggt tattaaaaga actaacacaa aagaaaactc
     1501 acaaggcaaa tatagagatt agccttgatg aatttaagtt catgttaatg cttgaaaata
     1561 actaccatga gtttaaaagg cttaaccaat gggttttgaa accaataagt aaagatttaa
     1621 acacttacag caatatgaaa ttggtggttg ataagcgagg ccgcccgact gatacgttga
     1681 ttttccaagt tgaactagat agacaaatgg atctcgtaac cgaacttgag aacaaccaga
     1741 taaaaatgaa tggtgacaaa ataccaacaa ccattacatc agattcctac ctacataacg
     1801 gactaagaaa aacactacac gatgctttaa ctgcaaaaat tcagctcacc agttttgagg
     1861 caaaattttt gagtgacatg caaagtaagt atgatctcaa tggttcgttc tcatggctca
     1921 cgcaaaaaca acgaaccaca ctagagaaca tactggctaa atacggaagg atctgaggtt
     1981 cttatggctc ttgtatctat cagtgaagca tcaagactaa caaacaaaag tagaacaact
     2041 gttcaccgtt acatatcaaa gggaaaactg tccatatgca cagatgaaaa cggtgtaaaa
     2101 aagatagata catcagagct tttacgagtt tttggtgcat tcaaagctgt tcaccatgaa
     2161 cagatcgaca atgtaacaga tgaacagcat gtaacaccta atagaacagg tgaaaccagt
     2221 aaaacaaagc aactagaaca tgaaattgaa cacctgagac aacttgttac agctcaacag
     2281 tcacacatag acagcctgaa acaggcgatg ctgcttatcg aatcaaagct gccgacaaca
     2341 cgggagccag tgacgcctcc cgtggggaaa aaatcatggc aattctggaa gaaatagcgc
     2401 tttcagccgg caaaccggct gaagccggat ctgcgattct gataacaaac tagcaacacc
     2461 agaacagccc gtttgcgggc agcaaaaccc gtaccctagg tataaacgca gaaaggccca
     2521 cccgaaggtg agccagtgtg actctagtag agagcgttca ccgacaaaca acagataaaa
     2581 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctgga gatccttact
     2641 cgagtttgga tccttaagca ccggtggagt gacgaccttc agcacgttcg tactgttcaa
     2701 cgatggtgta gtcttcgttg tgggaggtga tgtccagttt gatgtcggtt ttgtaagcac
     2761 ccggcagctg aaccggtttt ttagccatgt aggtggtttt aacttcagcg tcgtagtgac
     2821 caccgtcttt cagtttcaga cgcattttga tttcaccttt cagagcaccg tcttccgggt
     2881 acatacgttc ggtggaagct tcccaaccca tggttttttt ctgcataacc ggaccgtcgg
     2941 acgggaagtt ggtaccacgc agtttaactt tgtagatgaa ctcaccgtct tgcagggagg
     3001 agtcctgggt aacggtaaca acaccaccgt cttcgaagtt cataacacgt tcccatttga
     3061 aaccttccgg gaaggacagt ttcaggtagt ccgggatgtc agccgggtgt ttaacgtaag
     3121 ctttggaacc gtactggaac tgcggggaca ggatgtccca agcgaacggc agcggaccac
     3181 ctttggtaac tttcagttta gcggtctggg taccttcgta cggacgacct tcaccttcac
     3241 cttcgatttc gaactcgtga ccgttaacgg aaccttccat acgaactttg aaacgcatga
     3301 actctttgat aacgtcttcg ctactcgcca tatgtatatc tccttcttaa aagatctttt
     3361 gaattcccaa aaaaacgggt atggagaaac agtagagagt tgcgataaaa agcgtcaggt
     3421 agaatccgct aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa
     3481 ataatcaatg tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat
     3541 ggctttggtc ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag
     3601 cgagaagagc cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt
     3661 ttcttctctg aatggcggga gtatgaaaag tatggctgaa gcgcaaaatg atcccctgct
     3721 gccgggatac tcgtttaatg cccatctggt ggcgggttta acgccgattg aggccaacgg
     3781 ttatctcgat ttttttatcg accgaccgct gggaatgaaa ggttatattc tcaatctcac
     3841 cattcgcggt cagggggtgg tgaaaaatca gggacgagaa tttgtttgcc gaccgggtga
     3901 tattttgctg ttcccgccag gagagattca tcactacggt cgtcatccgg aggctcgcga
     3961 atggtatcac cagtgggttt actttcgtcc gcgcgcctac tggcatgaat ggcttaactg
     4021 gccgtcaata tttgccaata cggggttctt tcgcccggat gaagcgcacc agccgcattt
     4081 cagcgacctg tttgggcaaa tcattaacgc cgggcaaggg gaagggcgct attcggagct
     4141 gctggcgata aatctgcttg agcaattgtt actgcggcgc atggaagcga ttaacgagtc
     4201 gctccatcca ccgatggata atcgggtacg cgaggcttgt cagtacatca gcgatcacct
     4261 ggcagacagc aattttgata tcgccagcgt cgcacagcat gtttgcttgt cgccgtcgcg
     4321 tctgtcacat cttttccgcc agcagttagg gattagcgtc ttaagctggc gcgaggacca
     4381 acgtatcagc caggcgaagc tgcttttgag caccacccgg atgcctatcg ccaccgtcgg
     4441 tcgcaatgtt ggttttgacg atcaactcta tttctcgcgg gtatttaaaa aatgcaccgg
     4501 ggccagcccg agcgagttcc gtgccggttg tgaagaaaaa gtgaatgatg tagccgtcaa
     4561 gttgtcataa gacgtcggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct
     4621 gcaaagtaaa ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcatgat
     4681 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4741 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4801 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4861 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4921 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4981 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5041 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5101 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5161 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5221 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5281 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5341 gccggctggg tgtggcggac cgctatca
//
