LOCUS       sequence-493149-        3530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 expression vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3530)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3530)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSC101TSa2_Blue
FEATURES             Location/Qualifiers
     source          1..3530
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..73
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        127..145
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     127..144
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    152..208
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(210..229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(211..229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(236..253)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(236..252)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(245..267)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(461..480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        565..669
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(670..738,739..1530)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(888..907)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2152..2374
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2422..3372
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
ORIGIN
        1 cgtctcagtc aggagagaga cccctgttga caattaatca tcggcatagt atatcggcat
       61 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgacc atgattacgc
      121 caagctattt aggtgacact atagaatact caagcttgca tgcctgcagg tcgactctag
      181 aggatccccg ggtaccgagc tcgaattcgc cctatagtga gtcgtattac aattcactgg
      241 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      301 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      361 cccaacagtt gcgcagcctg aatggcgaat gacgcctgat gcggtatttt ctccttacgc
      421 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
      481 catagggtct ctcgcttgag acgaagcttg acacggttat ccacagaatc aggggataca
      541 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
      601 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
      661 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
      721 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
      781 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
      841 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
      901 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
      961 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1021 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1081 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     1141 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     1201 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     1261 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     1321 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     1381 cgtggttctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     1441 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     1501 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     1561 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1621 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgtt
     1681 gatgataccg ctgccttact gggtgcatta gccagtctga atgacctgtc acgggataat
     1741 ccgaagtggt cagactggaa aatcagaggg caggaactgc tgaacagcaa aaagtcagat
     1801 agcaccacat agcagacccg ccataaaacg ccctgagaag cccgtgacgg gcttttcttg
     1861 tattatgggt agtttccttg catgaatcca taaaaggcgc ctgtagtgcc atttaccccc
     1921 attcactgcc agagccgtga gcgcagcgaa ctgaatgtca cgaaaaagac agcgactcag
     1981 gtgcctgatg gtcggagaca aaaggaatat tcagcgattt gcccgattgc ggccgcaacc
     2041 gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca
     2101 aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac
     2161 acagggctgg gatctattct ttttatcttt ttttattctt tctttattct ataaattata
     2221 accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta
     2281 gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa
     2341 aggaaaagga ctagtaatta tcattgacta gcccatctca attggtatag tgattaaaat
     2401 cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg
     2461 attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta
     2521 ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac
     2581 caatacgttc agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca
     2641 accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttgag
     2701 attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt
     2761 gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat
     2821 gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta
     2881 acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg
     2941 ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca
     3001 ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc
     3061 ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa
     3121 cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat
     3181 tcctacctac ataacggact aagaaaaaca ctacacgatg ctttaactgc aaaaattcag
     3241 ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt
     3301 tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac
     3361 ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa
     3421 caaaagtaga acaactgttc accgttacat atcaaaggga aaactgtcca tacccatgga
     3481 tttaacgcga attttaacaa aatattaacg cttacaattt taacaagctt
//
