LOCUS       sequence-212797-        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
FEATURES             Location/Qualifiers
     source          1..5368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      169..263
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1025..1975
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTWKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2506..2533)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2549..2620)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2653..3330)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     promoter        complement(3380..3664)
                     /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(3438..3457)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3691..4569
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     CDS             join(4712..5368,1..138)
                     /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(4766..4785)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
       61 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
      121 cttgacgagt tcttctgagc gggactctgg ggttcgagag ctcgcttgga ctcctgttga
      181 tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc ggttgccgcc
      241 gggcgttttt tattggtgag aatccaagca ctagggacag taagacgggt aagcctgttg
      301 atgataccgc tgccttactg ggtgcattag ccagtctgaa tgacctgtca cgggataatc
      361 cgaagtggtc agactggaaa atcagagggc aggaactgct gaacagcaaa aagtcagata
      421 gcaccacata gcagacccgc cataaaacgc cctgagaagc ccgtgacggg cttttcttgt
      481 attatgggta gtttccttgc atgaatccat aaaaggcgcc tgtagtgcca tttaccccca
      541 ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg
      601 tgcctgatgg tcggagacaa aaggaatatt cagcgatttg cccgagcttg cgagggtgct
      661 acttaagcct ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat
      721 ccttttgtaa tactgcggaa ctgactaaag tagtgagtta tacacagggc tgggatctat
      781 tctttttatc tttttttatt ctttctttat tctataaatt ataaccactt gaatataaac
      841 aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt
      901 tccagcaaag gtctagcaga atttacagat acccagatca cccgggaaaa ggactagtaa
      961 ttatcattga ctagcccatc tcaattggta tagtgattaa aatcacctag accaattgag
     1021 atgtatgtct gaattagttg ttttcaaagc aaatgaacta gcgattagtc gctatgactt
     1081 aacggagcat gaaaccaagc taattttatg ctgtgtggca ctactcaacc ccacgattga
     1141 aaaccctaca tggaaagaac ggacggtatc gttcacttat aaccaatacg ctcagatgat
     1201 gaacatcagt agggaaaatg cttatggtgt attagctaaa gcaaccagag agctgatgac
     1261 gagaactgtg gaaatcagga atcctttggt taaaggcttt gagattttcc agtggacaaa
     1321 ctatgccaag ttctcaagcg aaaaattaga attagttttt agtgaagaga tattgcctta
     1381 tcttttccag ttaaaaaaat tcataaaata taatctggaa catgttaagt cttttgaaaa
     1441 caaatactct atgaggattt atgagtggtt attaaaagaa ctaacacaaa agaaaactca
     1501 caaggcaaat atagagatta gccttgatga atttaagttc atgttaatgc ttgaaaataa
     1561 ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta aagatttaaa
     1621 cacttacagc aatatgaaat tggtggttga taagcgaggc cgcccgactg atacgttgat
     1681 tttccaagtt gaactagata gacaaatgga tctcgtaacc gaacttgaga acaaccagat
     1741 aaaaatgaat ggtgacaaaa taccaacaac cattacatca gattcctacc tacataacgg
     1801 actaagaaaa acactacacg atgctttaac tgcaaaaatt cagctcacca gttttgaggc
     1861 aaaatttttg agtgacatgc aaagtaagta tgatctcaat ggttcgttct catggctcac
     1921 gcaaaaacaa cgaaccacac tagagaacat actggctaaa tacggaagga tctgaggttc
     1981 ttatggctct tgtatctatc agtgaagcat caagactaac aaacaaaagt agaacaactg
     2041 ttcaccgtta catatcaaag ggaaaactgt ccatatgcac agatgaaaac ggtgtaaaaa
     2101 agatagatac atcagagctt ttacgagttt ttggtgcatt caaagctgtt caccatgaac
     2161 agatcgacaa tgtaacagat gaacagcatg taacacctaa tagaacaggt gaaaccagta
     2221 aaacaaagca actagaacat gaaattgaac acctgagaca acttgttaca gctcaacagt
     2281 cacacataga cagcctgaaa caggcgatgc tgcttatcga atcaaagctg ccgacaacac
     2341 gggagccagt gacgcctccc gtggggaaaa aatcatggca attctggaag aaatagcgct
     2401 ttcagccggc aaaccggctg aagccggatc tgcgattctg ataacaaact agcaacacca
     2461 gaacagcccg tttgcgggca gcaaaacccg taccctaggt ataaacgcag aaaggcccac
     2521 ccgaaggtga gccagtgtga ctctagtaga gagcgttcac cgacaaacaa cagataaaac
     2581 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggag atccttactc
     2641 gagtttggat ccttaagcac cggtggagtg acgaccttca gcacgttcgt actgttcaac
     2701 gatggtgtag tcttcgttgt gggaggtgat gtccagtttg atgtcggttt tgtaagcacc
     2761 cggcagctga accggttttt tagccatgta ggtggtttta acttcagcgt cgtagtgacc
     2821 accgtctttc agtttcagac gcattttgat ttcacctttc agagcaccgt cttccgggta
     2881 catacgttcg gtggaagctt cccaacccat ggtttttttc tgcataaccg gaccgtcgga
     2941 cgggaagttg gtaccacgca gtttaacttt gtagatgaac tcaccgtctt gcagggagga
     3001 gtcctgggta acggtaacaa caccaccgtc ttcgaagttc ataacacgtt cccatttgaa
     3061 accttccggg aaggacagtt tcaggtagtc cgggatgtca gccgggtgtt taacgtaagc
     3121 tttggaaccg tactggaact gcggggacag gatgtcccaa gcgaacggca gcggaccacc
     3181 tttggtaact ttcagtttag cggtctgggt accttcgtac ggacgacctt caccttcacc
     3241 ttcgatttcg aactcgtgac cgttaacgga accttccata cgaactttga aacgcatgaa
     3301 ctctttgata acgtcttcgc tactcgccat atgtatatct ccttcttaaa agatcttttg
     3361 aattcccaaa aaaacgggta tggagaaaca gtagagagtt gcgataaaaa gcgtcaggta
     3421 gaatccgcta atcttatgga taaaaatgct atggcatagc aaagtgtgac gccgtgcaaa
     3481 taatcaatgt ggacttttct gccgtgatta tagacacttt tgttacgcgt ttttgtcatg
     3541 gctttggtcc cgctttgtta cagaatgctt ttaataagcg gggttaccgg tttggttagc
     3601 gagaagagcc agtaaaagac gcagtgacgg caatgtctga tgcaatatgg acaattggtt
     3661 tcttctctga atggcgggag tatgaaaagt atggctgaag cgcaaaatga tcccctgctg
     3721 ccgggatact cgtttaatgc ccatctggtg gcgggtttaa cgccgattga ggccaacggt
     3781 tatctcgatt tttttatcga ccgaccgctg ggaatgaaag gttatattct caatctcacc
     3841 attcgcggtc agggggtggt gaaaaatcag ggacgagaat ttgtttgccg accgggtgat
     3901 attttgctgt tcccgccagg agagattcat cactacggtc gtcatccgga ggctcgcgaa
     3961 tggtatcacc agtgggttta ctttcgtccg cgcgcctact ggcatgaatg gcttaactgg
     4021 ccgtcaatat ttgccaatac ggggttcttt cgcccggatg aagcgcacca gccgcatttc
     4081 agcgacctgt ttgggcaaat cattaacgcc gggcaagggg aagggcgcta ttcggagctg
     4141 ctggcgataa atctgcttga gcaattgtta ctgcggcgca tggaagcgat taacgagtcg
     4201 ctccatccac cgatggataa tcgggtacgc gaggcttgtc agtacatcag cgatcacctg
     4261 gcagacagca attttgatat cgccagcgtc gcacagcatg tttgcttgtc gccgtcgcgt
     4321 ctgtcacatc ttttccgcca gcagttaggg attagcgtct taagctggcg cgaggaccaa
     4381 cgtatcagcc aggcgaagct gcttttgagc accacccgga tgcctatcgc caccgtcggt
     4441 cgcaatgttg gttttgacga tcaactctat ttctcgcggg tatttaaaaa atgcaccggg
     4501 gccagcccga gcgagttccg tgccggttgt gaagaaaaag tgaatgatgt agccgtcaag
     4561 ttgtcataag acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg
     4621 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcatgatc
     4681 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4741 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4801 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4861 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4921 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4981 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5041 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     5101 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     5161 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     5221 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     5281 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     5341 ccggctgggt gtggcggacc gctatcag
//
