LOCUS       sequence-212791-        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_E93G
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(176..270)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(301..1095)
                     /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(412..431)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1022..1041
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1238..2116)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2143..2427
                     /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     2350..2369
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2477..3154
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3187..3258
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3274..3301
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3832..4782)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gggtctgcta tgtggtgcta tctgactttt tgctgttcag cagttcctgc cctctgattt
       61 tccagtctga ccacttcgga ttatcccgtg acaggtcatt cagactggct aatgcaccca
      121 gtaaggcagc ggtatcatca acaggcttac ccgtcttact gtccctagtg cttggattct
      181 caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg
      241 aggtcattac tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcccgc
      301 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
      361 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
      421 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
      481 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
      541 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
      601 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
      661 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
      721 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
      781 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
      841 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
      901 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
      961 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     1021 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     1081 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc atgatcccct
     1141 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
     1201 cttaccagag ggcgccccag ctggcaattc cgacgtctta tgacaacttg acggctacat
     1261 cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt
     1321 taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga
     1381 taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc
     1441 agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca
     1501 agcaaacatg ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga
     1561 tgtactgaca agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg
     1621 cttccatgcg ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc
     1681 gcccttcccc ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt
     1741 gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt
     1801 catgccagta ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg
     1861 gatgacgacc gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc
     1921 aaacaaattc tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa
     1981 tataaccttt cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa
     2041 tcggcgttaa acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat
     2101 tttgcgcttc agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat
     2161 attgcatcag acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg
     2221 taaccccgct tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc
     2281 gtaacaaaag tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca
     2341 cactttgcta tgccatagca tttttatcca taagattagc ggattctacc tgacgctttt
     2401 tatcgcaact ctctactgtt tctccatacc cgtttttttg ggaattcaaa agatctttta
     2461 agaaggagat atacatatgg cgagtagcga agacgttatc aaagagttca tgcgtttcaa
     2521 agttcgtatg gaaggttccg ttaacggtca cgagttcgaa atcgaaggtg aaggtgaagg
     2581 tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt accaaaggtg gtccgctgcc
     2641 gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt tccaaagctt acgttaaaca
     2701 cccggctgac atcccggact acctgaaact gtccttcccg gaaggtttca aatgggaacg
     2761 tgttatgaac ttcgaagacg gtggtgttgt taccgttacc caggactcct ccctgcaaga
     2821 cggtgagttc atctacaaag ttaaactgcg tggtaccaac ttcccgtccg acggtccggt
     2881 tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa cgtatgtacc cggaagacgg
     2941 tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa gacggtggtc actacgacgc
     3001 tgaagttaaa accacctaca tggctaaaaa accggttcag ctgccgggtg cttacaaaac
     3061 cgacatcaaa ctggacatca cctcccacaa cgaagactac accatcgttg aacagtacga
     3121 acgtgctgaa ggtcgtcact ccaccggtgc ttaaggatcc aaactcgagt aaggatctcc
     3181 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3241 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     3301 gtttatacct agggtacggg ttttgctgcc cgcaaacggg ctgttctggt gttgctagtt
     3361 tgttatcaga atcgcagatc cggcttcagc cggtttgccg gctgaaagcg ctatttcttc
     3421 cagaattgcc atgatttttt ccccacggga ggcgtcactg gctcccgtgt tgtcggcagc
     3481 tttgattcga taagcagcat cgcctgtttc aggctgtcta tgtgtgactg ttgagctgta
     3541 acaagttgtc tcaggtgttc aatttcatgt tctagttgct ttgttttact ggtttcacct
     3601 gttctattag gtgttacatg ctgttcatct gttacattgt cgatctgttc atggtgaaca
     3661 gctttgaatg caccaaaaac tcgtaaaagc tctgatgtat ctatcttttt tacaccgttt
     3721 tcatctgtgc atatggacag ttttcccttt gatatgtaac ggtgaacagt tgttctactt
     3781 ttgtttgtta gtcttgatgc ttcactgata gatacaagag ccataagaac ctcagatcct
     3841 tccgtattta gccagtatgt tctctagtgt ggttcgttgt ttttgcgtga gccatgagaa
     3901 cgaaccattg agatcatact tactttgcat gtcactcaaa aattttgcct caaaactggt
     3961 gagctgaatt tttgcagtta aagcatcgtg tagtgttttt cttagtccgt tatgtaggta
     4021 ggaatctgat gtaatggttg ttggtatttt gtcaccattc atttttatct ggttgttctc
     4081 aagttcggtt acgagatcca tttgtctatc tagttcaact tggaaaatca acgtatcagt
     4141 cgggcggcct cgcttatcaa ccaccaattt catattgctg taagtgttta aatctttact
     4201 tattggtttc aaaacccatt ggttaagcct tttaaactca tggtagttat tttcaagcat
     4261 taacatgaac ttaaattcat caaggctaat ctctatattt gccttgtgag ttttcttttg
     4321 tgttagttct tttaataacc actcataaat cctcatagag tatttgtttt caaaagactt
     4381 aacatgttcc agattatatt ttatgaattt ttttaactgg aaaagataag gcaatatctc
     4441 ttcactaaaa actaattcta atttttcgct tgagaacttg gcatagtttg tccactggaa
     4501 aatcccaaag cctttaacca aaggattcct gatttccaca gttctcgtca tcagctctct
     4561 ggttgcttta gctaatacac cataagcatt ttccctactg atgttcatca tctgagcgta
     4621 ttggttataa gtgaacgata ccgtccgttc tttccttgta gggttttcaa tcgtggggtt
     4681 gagtagtgcc acacagcata aaattagctt ggtttcatgc tccgttaagt catagcgact
     4741 aatcgctagt tcatttgctt tgaaaacaac taattcagac atacatctca attggtctag
     4801 gtgattttaa tcactatacc aattgagatg ggctagtcaa tgataattac tagtcctttt
     4861 cccgggtgat ctgggtatct gtaaattctg ctagaccttt gctggaaaac ttgtaaattc
     4921 tgctagaccc tctgtaaatt ccgctagacc tttgtgtgtt ttttttgttt atattcaagt
     4981 ggttataatt tatagaataa agaaagaata aaaaaagata aaaagaatag atcccagccc
     5041 tgtgtataac tcactacttt agtcagttcc gcagtattac aaaaggatgt cgcaaacgct
     5101 gtttgctcct ctacaaaaca gaccttaaaa ccctaaaggc ttaagtagca ccctcgcaag
     5161 ctcgggcaaa tcgctgaata ttccttttgt ctccgaccat caggcacctg agtcgctgtc
     5221 tttttcgtga cattcagttc gctgcgctca cggctctggc agtgaatggg ggtaaatggc
     5281 actacaggcg ccttttatgg attcatgcaa ggaaactacc cataatacaa gaaaagcccg
     5341 tcacgggctt ctcagggcgt tttatggc
//
