LOCUS       sequence-212798-        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_R43W_E93G
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      87..181
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             943..1893
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTWKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2424..2451)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2467..2538)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2571..3248)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3298..3582)
                     /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(3356..3375)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3609..4487
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4630..5368,1..56)
                     /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(4684..4703)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5294..5313
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
       61 gactctgggg ttcgagagct cgcttggact cctgttgata gatccagtaa tgacctcaga
      121 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
      181 tccaagcact agggacagta agacgggtaa gcctgttgat gataccgctg ccttactggg
      241 tgcattagcc agtctgaatg acctgtcacg ggataatccg aagtggtcag actggaaaat
      301 cagagggcag gaactgctga acagcaaaaa gtcagatagc accacatagc agacccgcca
      361 taaaacgccc tgagaagccc gtgacgggct tttcttgtat tatgggtagt ttccttgcat
      421 gaatccataa aaggcgcctg tagtgccatt tacccccatt cactgccaga gccgtgagcg
      481 cagcgaactg aatgtcacga aaaagacagc gactcaggtg cctgatggtc ggagacaaaa
      541 ggaatattca gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa
      601 ggtctgtttt gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact
      661 gactaaagta gtgagttata cacagggctg ggatctattc tttttatctt tttttattct
      721 ttctttattc tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta
      781 gcggaattta cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat
      841 ttacagatac ccagatcacc cgggaaaagg actagtaatt atcattgact agcccatctc
      901 aattggtata gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt
      961 ttcaaagcaa atgaactagc gattagtcgc tatgacttaa cggagcatga aaccaagcta
     1021 attttatgct gtgtggcact actcaacccc acgattgaaa accctacatg gaaagaacgg
     1081 acggtatcgt tcacttataa ccaatacgct cagatgatga acatcagtag ggaaaatgct
     1141 tatggtgtat tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat
     1201 cctttggtta aaggctttgg gattttccag tggacaaact atgccaagtt ctcaagcgaa
     1261 aaattagaat tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc
     1321 ataaaatata atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat
     1381 gagtggttat taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc
     1441 cttgatgaat ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt
     1501 aaccaatggg ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg
     1561 gtggttgata agcgaggccg cccgactgat acgttgattt tccaagttga actagataga
     1621 caaatggatc tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata
     1681 ccaacaacca ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat
     1741 gctttaactg caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa
     1801 agtaagtatg atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta
     1861 gagaacatac tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag
     1921 tgaagcatca agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg
     1981 aaaactgtcc atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt
     2041 acgagttttt ggtgcattca aagctgttca ccatgaacag atcgacaatg taacagatga
     2101 acagcatgta acacctaata gaacaggtga aaccagtaaa acaaagcaac tagaacatga
     2161 aattgaacac ctgagacaac ttgttacagc tcaacagtca cacatagaca gcctgaaaca
     2221 ggcgatgctg cttatcgaat caaagctgcc gacaacacgg gagccagtga cgcctcccgt
     2281 ggggaaaaaa tcatggcaat tctggaagaa atagcgcttt cagccggcaa accggctgaa
     2341 gccggatctg cgattctgat aacaaactag caacaccaga acagcccgtt tgcgggcagc
     2401 aaaacccgta ccctaggtat aaacgcagaa aggcccaccc gaaggtgagc cagtgtgact
     2461 ctagtagaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     2521 agcctttcgt tttatttgat gcctggagat ccttactcga gtttggatcc ttaagcaccg
     2581 gtggagtgac gaccttcagc acgttcgtac tgttcaacga tggtgtagtc ttcgttgtgg
     2641 gaggtgatgt ccagtttgat gtcggttttg taagcacccg gcagctgaac cggtttttta
     2701 gccatgtagg tggttttaac ttcagcgtcg tagtgaccac cgtctttcag tttcagacgc
     2761 attttgattt cacctttcag agcaccgtct tccgggtaca tacgttcggt ggaagcttcc
     2821 caacccatgg tttttttctg cataaccgga ccgtcggacg ggaagttggt accacgcagt
     2881 ttaactttgt agatgaactc accgtcttgc agggaggagt cctgggtaac ggtaacaaca
     2941 ccaccgtctt cgaagttcat aacacgttcc catttgaaac cttccgggaa ggacagtttc
     3001 aggtagtccg ggatgtcagc cgggtgttta acgtaagctt tggaaccgta ctggaactgc
     3061 ggggacagga tgtcccaagc gaacggcagc ggaccacctt tggtaacttt cagtttagcg
     3121 gtctgggtac cttcgtacgg acgaccttca ccttcacctt ctatttcgaa ctcgtgaccg
     3181 ttaacggaac cttccatacg aactttgaaa cgcatgaact ctttgataac gtcttcgcta
     3241 ctcgccatat gtatatctcc ttcttaaaag atcttttgaa ttcccaaaaa aacgggtatg
     3301 gagaaacagt agagagttgc gataaaaagc gtcaggtaga atccgctaat cttatggata
     3361 aaaatgctat ggcatagcaa agtgtgacgc cgtgcaaata atcaatgtgg acttttctgc
     3421 cgtgattata gacacttttg ttacgcgttt ttgtcatggc tttggtcccg ctttgttaca
     3481 gaatgctttt aataagcggg gttaccggtt tggttagcga gaagagccag taaaagacgc
     3541 agtgacggca atgtctgatg caatatggac aattggtttc ttctctgaat ggcgggagta
     3601 tgaaaagtat ggctgaagcg caaaatgatc ccctgctgcc gggatactcg tttaatgccc
     3661 atctggtggc gggtttaacg ccgattgagg ccaacggtta tctcgatttt tttatcgacc
     3721 gaccgctggg aatgaaaggt tatattctca atctcaccat tcgcggtcag ggggtggtga
     3781 aaaatcaggg acgagaattt gtttgccgac cgggtgatat tttgctgttc ccgccaggag
     3841 agattcatca ctacggtcgt catccggagg ctcgcgaatg gtatcaccag tgggtttact
     3901 ttcgtccgcg cgcctactgg catgaatggc ttaactggcc gtcaatattt gccaatacgg
     3961 ggttctttcg cccggatgaa gcgcaccagc cgcatttcag cgacctgttt gggcaaatca
     4021 ttaacgccgg gcaaggggaa gggcgctatt cggagctgct ggcgataaat ctgcttgagc
     4081 aattgttact gcggcgcatg gaagcgatta acgagtcgct ccatccaccg atggataatc
     4141 gggtacgcga ggcttgtcag tacatcagcg atcacctggc agacagcaat tttgatatcg
     4201 ccagcgtcgc acagcatgtt tgcttgtcgc cgtcgcgtct gtcacatctt ttccgccagc
     4261 agttagggat tagcgtctta agctggcgcg aggaccaacg tatcagccag gcgaagctgc
     4321 ttttgagcac cacccggatg cctatcgcca ccgtcggtcg caatgttggt tttgacgatc
     4381 aactctattt ctcgcgggta tttaaaaaat gcaccggggc cagcccgagc gagttccgtg
     4441 ccggttgtga agaaaaagtg aatgatgtag ccgtcaagtt gtcataagac gtcggaattg
     4501 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     4561 ttgccgccaa ggatctgatg gcgcagggga tcatgatctg atcaagagac aggatgagga
     4621 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4681 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4741 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4801 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4861 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4921 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4981 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5041 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5101 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5161 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5221 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5281 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5341 gaccgcttcc tcgtgcttta cggtatcg
//
