LOCUS       sequence-212794-        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_R46Q
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(217..311)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(342..1136)
                     /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(453..472)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1063..1082
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1279..2157)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2184..2468
                     /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     2391..2410
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2518..3195
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3228..3299
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3315..3342
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3873..4823)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     QTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 caagaaaagc ccgtcacggg cttctcaggg cgttttatgg cgggtctgct atgtggtgct
       61 atctgacttt ttgctgttca gcagttcctg ccctctgatt ttccagtctg accacttcgg
      121 attatcccgt gacaggtcat tcagactggc taatgcaccc agtaaggcag cggtatcatc
      181 aacaggctta cccgtcttac tgtccctagt gcttggattc tcaccaataa aaaacgcccg
      241 gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat
      301 caacaggagt ccaagcgagc tctcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
      361 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
      421 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
      481 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
      541 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
      601 ggcatgcgcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
      661 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
      721 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
      781 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
      841 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
      901 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcctgcagt
      961 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     1021 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     1081 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     1141 aacgatcctc atcctgtctc ttgatcagat catgatcccc tgcgccatca gatccttggc
     1201 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     1261 gctggcaatt ccgacgtctt atgacaactt gacggctaca tcattcactt tttcttcaca
     1321 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
     1381 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     1441 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     1501 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     1561 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     1621 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
     1681 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     1741 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     1801 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     1861 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     1921 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
     1981 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     2041 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     2101 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     2161 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
     2221 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     2281 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     2341 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     2401 atttttatcc ataagattag cggattctac ctgacgcttt ttatcgcaac tctctactgt
     2461 ttctccatac ccgttttttt gggaattcaa aagatctttt aagaaggaga tatacatatg
     2521 gcgagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc
     2581 gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc
     2641 cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg
     2701 tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac
     2761 tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac
     2821 ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa
     2881 gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg
     2941 ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc
     3001 aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac
     3061 atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc
     3121 acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac
     3181 tccaccggtg cttaaggatc caaactcgag taaggatctc caggcatcaa ataaaacgaa
     3241 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     3301 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatacc tagggtacgg
     3361 gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag aatcgcagat
     3421 ccggcttcag ccggtttgcc ggctgaaagc gctatttctt ccagaattgc catgattttt
     3481 tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg ataagcagca
     3541 tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt
     3601 caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta ggtgttacat
     3661 gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttgaat gcaccaaaaa
     3721 ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg catatggaca
     3781 gttttccctt tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg
     3841 cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg
     3901 ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac
     3961 ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt
     4021 aaagcatcgt gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt
     4081 gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc
     4141 atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca
     4201 accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat
     4261 tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca
     4321 tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac
     4381 cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat
     4441 tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct
     4501 aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc
     4561 aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca
     4621 ccataagcat tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat
     4681 accgtctgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat
     4741 aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct
     4801 ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac
     4861 caattgagat gggctagtca atgataatta ctagtccttt tcccgggtga tctgggtatc
     4921 tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat
     4981 tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata
     5041 aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt
     5101 tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac
     5161 agaccttaaa accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat
     5221 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
     5281 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
     5341 gattcatgca aggaaactac ccataata
//
