LOCUS       sequence-493159-        3930 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 3930)
  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  JOURNAL   Unpublished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 3930)
  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            "Unpublished"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: pSC101TSo1_Red
FEATURES             Location/Qualifiers
     source          1..3930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..69
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      809..880
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        965..1069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1070..1138,1139..1930)
                     /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(1288..1307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2552..2774
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2822..3772
                     /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 ggtctcagga gagagacgcc tgttgacaat taatcatcgg catagtatat cggcatagta
       61 taatacgacc aaagaaacaa aatgaggagg tactgaaatg gtgagcaaag gcgaagcggt
      121 gattaaagaa tttatgcgct ttaaagtgca tatggaaggc agcatgaacg gccatgaatt
      181 tgaaattgaa ggcgaaggcg aaggccgccc gtatgaaggc acccagaccg cgaaactgaa
      241 agtgaccaaa ggcggcccgc tgccgtttag ctgggatatt ctgagcccgc agtttatgta
      301 tggcagccgc gcgtttatta aacatccggc ggatattccg gattattata aacagagctt
      361 tccggaaggc tttaaatggg aacgcgtgat gaactttgaa gatggcggcg cggtgaccgt
      421 gacccaggat accagcctgg aagatggcac cctgatttat aaagtgaaac tgcgcggcac
      481 caactttccg ccggatggcc cggtgatgca gaaaaaaacc atgggctggg aagcgagcac
      541 cgaacgcctg tatccggaag atggcgtgct gaaaggcgat attaaaatgg cgctgcgcct
      601 gaaagatggc ggccgctatc tggcggattt taaaaccacc tataaagcga aaaaaccggt
      661 gcagatgccg ggcgcgtata acgtggatcg caaactggat attaccagcc ataacgaaga
      721 ttataccgtg gtggaacagt atgaacgcag cgaaggccgc catagcaccg gcggcatgga
      781 tgaactgtat aaataagctt ccaggcatca aataaaacga aaggctcagt cgaaagactg
      841 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc cgtctctcgc tgtcatgaga
      901 ccggatcttg acacggttat ccacagaatc aggggataca ggtggcactt ttcggggaaa
      961 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1021 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1081 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1141 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1201 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1261 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1321 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1381 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1441 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1501 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1561 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1621 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1681 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1741 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggttctc gcggtatcat
     1801 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1861 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1921 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1981 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2041 ttaacgtgag ttttcgttcc actgagcgtc agaccccgtt gatgataccg ctgccttact
     2101 gggtgcatta gccagtctga atgacctgtc acgggataat ccgaagtggt cagactggaa
     2161 aatcagaggg caggaactgc tgaacagcaa aaagtcagat agcaccacat agcagacccg
     2221 ccataaaacg ccctgagaag cccgtgacgg gcttttcttg tattatgggt agtttccttg
     2281 catgaatcca taaaaggcgc ctgtagtgcc atttaccccc attcactgcc agagccgtga
     2341 gcgcagcgaa ctgaatgtca cgaaaaagac agcgactcag gtgcctgatg gtcggagaca
     2401 aaaggaatat tcagcgattt gcccgattgc ggccgcaacc gagcttgcga gggtgctact
     2461 taagccttta gggttttaag gtctgttttg tagaggagca aacagcgttt gcgacatcct
     2521 tttgtaatac tgcggaactg actaaagtag tgagttatac acagggctgg gatctattct
     2581 ttttatcttt ttttattctt tctttattct ataaattata accacttgaa tataaacaaa
     2641 aaaaacacac aaaggtctag cggaatttac agagggtcta gcagaattta caagttttcc
     2701 agcaaaggtc tagcagaatt tacagatacc cacaactcaa aggaaaagga ctagtaatta
     2761 tcattgacta gcccatctca attggtatag tgattaaaat cacctagacc aattgagatg
     2821 tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg attagtcgct atgacttaac
     2881 ggagcatgaa accaagctaa ttttatgctg tgtggcacta ctcaacccca cgattgaaaa
     2941 ccctacaagg aaagaacgga cggtatcgtt cacttataac caatacgttc agatgatgaa
     3001 catcagtagg gaaaatgctt atggtgtatt agctaaagca accagagagc tgatgacgag
     3061 aactgtggaa atcaggaatc ctttggttaa aggctttgag attttccagt ggacaaacta
     3121 tgccaagttc tcaagcgaaa aattagaatt agtttttagt gaagagatat tgccttatct
     3181 tttccagtta aaaaaattca taaaatataa tctggaacat gttaagtctt ttgaaaacaa
     3241 atactctatg aggatttatg agtggttatt aaaagaacta acacaaaaga aaactcacaa
     3301 ggcaaatata gagattagcc ttgatgaatt taagttcatg ttaatgcttg aaaataacta
     3361 ccatgagttt aaaaggctta accaatgggt tttgaaacca ataagtaaag atttaaacac
     3421 ttacagcaat atgaaattgg tggttgataa gcgaggccgc ccgactgata cgttgatttt
     3481 ccaagttgaa ctagatagac aaatggatct cgtaaccgaa cttgagaaca accagataaa
     3541 aatgaatggt gacaaaatac caacaaccat tacatcagat tcctacctac ataacggact
     3601 aagaaaaaca ctacacgatg ctttaactgc aaaaattcag ctcaccagtt ttgaggcaaa
     3661 atttttgagt gacatgcaaa gtaagtatga tctcaatggt tcgttctcat ggctcacgca
     3721 aaaacaacga accacactag agaacatact ggctaaatac ggaaggatct gaggttctta
     3781 tggctcttgt atctatcagt gaagcatcaa gactaacaaa caaaagtaga acaactgttc
     3841 accgttacat atcaaaggga aaactgtcca tacccatgga tttaacgcga attttaacaa
     3901 aatattaacg cttacaattt taacggatcc
//
