LOCUS       sequence-493152-        3531 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 3531)
  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  JOURNAL   Unpublished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 3531)
  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            "Unpublished"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_Blue
FEATURES             Location/Qualifiers
     source          1..3531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..69
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        123..141
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     123..140
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    148..204
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(206..225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(207..225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(232..249)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(232..248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(241..263)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(457..476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        566..670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(671..739,740..1531)
                     /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(889..908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2153..2375
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2423..3373
                     /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 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgaccatga ttacgccaag
      121 ctatttaggt gacactatag aatactcaag cttgcatgcc tgcaggtcga ctctagagga
      181 tccccgggta ccgagctcga attcgcccta tagtgagtcg tattacaatt cactggccgt
      241 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      301 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      361 acagttgcgc agcctgaatg gcgaatgacg cctgatgcgg tattttctcc ttacgcatct
      421 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
      481 gcgtctctcg ctgtcatgag accggatctt gacacggtta tccacagaat caggggatac
      541 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      601 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      661 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      721 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      781 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      841 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      901 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      961 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1021 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1081 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1141 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1201 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1261 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1321 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1381 gcgtggttct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1441 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1501 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1561 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1621 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1681 tgatgatacc gctgccttac tgggtgcatt agccagtctg aatgacctgt cacgggataa
     1741 tccgaagtgg tcagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga
     1801 tagcaccaca tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt
     1861 gtattatggg tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc
     1921 cattcactgc cagagccgtg agcgcagcga actgaatgtc acgaaaaaga cagcgactca
     1981 ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt tgcccgattg cggccgcaac
     2041 cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt gtagaggagc
     2101 aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta gtgagttata
     2161 cacagggctg ggatctattc tttttatctt tttttattct ttctttattc tataaattat
     2221 aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta cagagggtct
     2281 agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac ccacaactca
     2341 aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata gtgattaaaa
     2401 tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa atgaactagc
     2461 gattagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct gtgtggcact
     2521 actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt tcacttataa
     2581 ccaatacgtt cagatgatga acatcagtag ggaaaatgct tatggtgtat tagctaaagc
     2641 aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta aaggctttga
     2701 gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat tagtttttag
     2761 tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata atctggaaca
     2821 tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat taaaagaact
     2881 aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat ttaagttcat
     2941 gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg ttttgaaacc
     3001 aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata agcgaggccg
     3061 cccgactgat acgttgattt tccaagttga actagataga caaatggatc tcgtaaccga
     3121 acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca ttacatcaga
     3181 ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg caaaaattca
     3241 gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg atctcaatgg
     3301 ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac tggctaaata
     3361 cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca agactaacaa
     3421 acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc atacccatgg
     3481 atttaacgcg aattttaaca aaatattaac gcttacaatt ttaacggatc c
//
