LOCUS       sequence-212800-        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_E115K
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(170..264)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(295..1089)
                     /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(406..425)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1016..1035
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1232..2110)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2137..2421
                     /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     2344..2363
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             2471..3148
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      3181..3252
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3268..3295
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3826..4776)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEKILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt
       61 ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg
      121 cagcggtatc atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa
      181 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
      241 ttactggatc tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa
      301 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
      361 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
      421 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
      481 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag
      541 atcctcgccg tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc
      601 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc
      661 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
      721 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
      781 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
      841 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
      901 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
      961 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     1021 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     1081 ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca
     1141 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     1201 agagggcgcc ccagctggca attccgacgt cttatgacaa cttgacggct acatcattca
     1261 ctttttcttc acaaccggca cggaactcgc tcgggctggc cccggtgcat tttttaaata
     1321 cccgcgagaa atagagttga tcgtcaaaac caacattgcg accgacggtg gcgataggca
     1381 tccgggtggt gctcaaaagc agcttcgcct ggctgatacg ttggtcctcg cgccagctta
     1441 agacgctaat ccctaactgc tggcggaaaa gatgtgacag acgcgacggc gacaagcaaa
     1501 catgctgtgc gacgctggcg atatcaaaat tgctgtctgc caggtgatcg ctgatgtact
     1561 gacaagcctc gcgtacccga ttatccatcg gtggatggag cgactcgtta atcgcttcca
     1621 tgcgccgcag taacaattgc tcaagcagat ttatcgccag cagctccgaa tagcgccctt
     1681 ccccttgccc ggcgttaatg atttgcccaa acaggtcgct gaaatgcggc tggtgcgctt
     1741 catccgggcg aaagaacccc gtattggcaa atattgacgg ccagttaagc cattcatgcc
     1801 agtaggcgcg cggacgaaag taaacccact ggtgatacca ttcgcgagcc tccggatgac
     1861 gaccgtagtg atgaatctct cctggcggga acagcaaaat atcacccggt cggcaaacaa
     1921 attctcgtcc ctgatttttc accaccccct gaccgcgaat ggtgagattg agaatataac
     1981 ctttcattcc cagcggtcgg tcgataaaaa aatcgagata accgttggcc tcaatcggcg
     2041 ttaaacccgc caccagatgg gcattaaacg agtatcccgg cagcagggga tcattttgcg
     2101 cttcagccat acttttcata ctcccgccat tcagagaaga aaccaattgt ccatattgca
     2161 tcagacattg ccgtcactgc gtcttttact ggctcttctc gctaaccaaa ccggtaaccc
     2221 cgcttattaa aagcattctg taacaaagcg ggaccaaagc catgacaaaa acgcgtaaca
     2281 aaagtgtcta taatcacggc agaaaagtcc acattgatta tttgcacggc gtcacacttt
     2341 gctatgccat agcattttta tccataagat tagcggattc tacctgacgc tttttatcgc
     2401 aactctctac tgtttctcca tacccgtttt tttgggaatt caaaagatct tttaagaagg
     2461 agatatacat atggcgagta gcgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg
     2521 tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc
     2581 gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc
     2641 ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc
     2701 tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat
     2761 gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga
     2821 gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca
     2881 gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct
     2941 gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt
     3001 taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat
     3061 caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc
     3121 tgaaggtcgt cactccaccg gtgcttaagg atccaaactc gagtaaggat ctccaggcat
     3181 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     3241 gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
     3301 acctagggta cgggttttgc tgcccgcaaa cgggctgttc tggtgttgct agtttgttat
     3361 cagaatcgca gatccggctt cagccggttt gccggctgaa agcgctattt cttccagaat
     3421 tgccatgatt ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat
     3481 tcgataagca gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt
     3541 tgtctcaggt gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta
     3601 ttaggtgtta catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagctttg
     3661 aatgcaccaa aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct
     3721 gtgcatatgg acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt
     3781 gttagtcttg atgcttcact gatagataca agagccataa gaacctcaga tccttccgta
     3841 tttagccagt atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc
     3901 attgagatca tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg
     3961 aatttttgca gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc
     4021 tgatgtaatg gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc
     4081 ggttacgaga tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg
     4141 gcctcgctta tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg
     4201 tttcaaaacc cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat
     4261 gaacttaaat tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag
     4321 ttcttttaat aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg
     4381 ttccagatta tattttatga atttttttaa ctggaaaaga taaggcaata tcttttcact
     4441 aaaaactaat tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc
     4501 aaagccttta accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc
     4561 tttagctaat acaccataag cattttccct actgatgttc atcatctgag cgtattggtt
     4621 ataagtgaac gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag
     4681 tgccacacag cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc
     4741 tagttcattt gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt
     4801 ttaatcacta taccaattga gatgggctag tcaatgataa ttactagtcc ttttcccggg
     4861 tgatctgggt atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag
     4921 accctctgta aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat
     4981 aatttataga ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta
     5041 taactcacta ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc
     5101 tcctctacaa aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg
     5161 caaatcgctg aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc
     5221 gtgacattca gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca
     5281 ggcgcctttt atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg
     5341 gcttctcagg gcgttttatg gcgggtct
//
