LOCUS       sequence-493157-        3929 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 3929)
  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  JOURNAL   Unpublished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 3929)
  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            "Unpublished"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_Red
FEATURES             Location/Qualifiers
     source          1..3929
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..73
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      813..884
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        964..1068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1069..1137,1138..1929)
                     /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(1287..1306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2551..2773
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2821..3771
                     /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 gaccaaagaa acaaaatgag gaggtactga aatggtgagc aaaggcgaag
      121 cggtgattaa agaatttatg cgctttaaag tgcatatgga aggcagcatg aacggccatg
      181 aatttgaaat tgaaggcgaa ggcgaaggcc gcccgtatga aggcacccag accgcgaaac
      241 tgaaagtgac caaaggcggc ccgctgccgt ttagctggga tattctgagc ccgcagttta
      301 tgtatggcag ccgcgcgttt attaaacatc cggcggatat tccggattat tataaacaga
      361 gctttccgga aggctttaaa tgggaacgcg tgatgaactt tgaagatggc ggcgcggtga
      421 ccgtgaccca ggataccagc ctggaagatg gcaccctgat ttataaagtg aaactgcgcg
      481 gcaccaactt tccgccggat ggcccggtga tgcagaaaaa aaccatgggc tgggaagcga
      541 gcaccgaacg cctgtatccg gaagatggcg tgctgaaagg cgatattaaa atggcgctgc
      601 gcctgaaaga tggcggccgc tatctggcgg attttaaaac cacctataaa gcgaaaaaac
      661 cggtgcagat gccgggcgcg tataacgtgg atcgcaaact ggatattacc agccataacg
      721 aagattatac cgtggtggaa cagtatgaac gcagcgaagg ccgccatagc accggcggca
      781 tggatgaact gtataaataa gcttccaggc atcaaataaa acgaaaggct cagtcgaaag
      841 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcggtctc tcgcttgaga
      901 cgaagcttga cacggttatc cacagaatca ggggatacag gtggcacttt tcggggaaat
      961 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     1021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     1081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     1141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     1261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     1321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     1381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     1441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1741 gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg cggtatcatt
     1801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     2041 taacgtgagt tttcgttcca ctgagcgtca gaccccgttg atgataccgc tgccttactg
     2101 ggtgcattag ccagtctgaa tgacctgtca cgggataatc cgaagtggtc agactggaaa
     2161 atcagagggc aggaactgct gaacagcaaa aagtcagata gcaccacata gcagacccgc
     2221 cataaaacgc cctgagaagc ccgtgacggg cttttcttgt attatgggta gtttccttgc
     2281 atgaatccat aaaaggcgcc tgtagtgcca tttaccccca ttcactgcca gagccgtgag
     2341 cgcagcgaac tgaatgtcac gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa
     2401 aaggaatatt cagcgatttg cccgattgcg gccgcaaccg agcttgcgag ggtgctactt
     2461 aagcctttag ggttttaagg tctgttttgt agaggagcaa acagcgtttg cgacatcctt
     2521 ttgtaatact gcggaactga ctaaagtagt gagttataca cagggctggg atctattctt
     2581 tttatctttt tttattcttt ctttattcta taaattataa ccacttgaat ataaacaaaa
     2641 aaaacacaca aaggtctagc ggaatttaca gagggtctag cagaatttac aagttttcca
     2701 gcaaaggtct agcagaattt acagataccc acaactcaaa ggaaaaggac tagtaattat
     2761 cattgactag cccatctcaa ttggtatagt gattaaaatc acctagacca attgagatgt
     2821 atgtctgaat tagttgtttt caaagcaaat gaactagcga ttagtcgcta tgacttaacg
     2881 gagcatgaaa ccaagctaat tttatgctgt gtggcactac tcaaccccac gattgaaaac
     2941 cctacaagga aagaacggac ggtatcgttc acttataacc aatacgttca gatgatgaac
     3001 atcagtaggg aaaatgctta tggtgtatta gctaaagcaa ccagagagct gatgacgaga
     3061 actgtggaaa tcaggaatcc tttggttaaa ggctttgaga ttttccagtg gacaaactat
     3121 gccaagttct caagcgaaaa attagaatta gtttttagtg aagagatatt gccttatctt
     3181 ttccagttaa aaaaattcat aaaatataat ctggaacatg ttaagtcttt tgaaaacaaa
     3241 tactctatga ggatttatga gtggttatta aaagaactaa cacaaaagaa aactcacaag
     3301 gcaaatatag agattagcct tgatgaattt aagttcatgt taatgcttga aaataactac
     3361 catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa taagtaaaga tttaaacact
     3421 tacagcaata tgaaattggt ggttgataag cgaggccgcc cgactgatac gttgattttc
     3481 caagttgaac tagatagaca aatggatctc gtaaccgaac ttgagaacaa ccagataaaa
     3541 atgaatggtg acaaaatacc aacaaccatt acatcagatt cctacctaca taacggacta
     3601 agaaaaacac tacacgatgc tttaactgca aaaattcagc tcaccagttt tgaggcaaaa
     3661 tttttgagtg acatgcaaag taagtatgat ctcaatggtt cgttctcatg gctcacgcaa
     3721 aaacaacgaa ccacactaga gaacatactg gctaaatacg gaaggatctg aggttcttat
     3781 ggctcttgta tctatcagtg aagcatcaag actaacaaac aaaagtagaa caactgttca
     3841 ccgttacata tcaaagggaa aactgtccat acccatggat ttaacgcgaa ttttaacaaa
     3901 atattaacgc ttacaatttt aacaagctt
//
