LOCUS       sequence-237131-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 1BC plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3488)
  AUTHORS   Taylor GM, Mordaka PM, Heap JT
  TITLE     Start-Stop Assembly: a functionally scarless DNA assembly system 
            optimized for metabolic engineering.
  JOURNAL   Nucleic Acids Res. 2018 Nov 20. pii: 5193345. doi: 
            10.1093/nar/gky1182.
  PUBMED    30462270
REFERENCE   2  (bases 1 to 3488)
  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: "Nucleic 
            Acids Res. 2018 Nov 20. pii: 5193345. doi: 10.1093/nar/gky1182."
COMMENT     Sequence Label: pStA1BC
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(92..178)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(581..598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(715..1260)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(795..814)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1372..1377,1378..1394,1395..1400)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1448..2638
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(1800..1819)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      2976..3070
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    3119..3140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3155..3160,3161..3178,3179..3185)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3193..3209
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3198..3220
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3217..3233
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3217..3233
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3229..3488,1..64)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    3246..3302
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3303..3320)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3303..3319)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3312..3334)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
       61 atagtttccg ctcttcagga aggtagagac catttgtcct actcaggaga gcgttcaccg
      121 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
      181 gcccgaaagg gcctcgtgat acgccaaaag ttggcccagg gcttcccggt atcaacaggg
      241 acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta ttcggcgcaa
      301 agtgtgtcgg gtgatgctgc caacttactg atttagtgta tgatggtgtt tttgaggtgc
      361 tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg gggtggtgcg
      421 taacggcaaa agcaccgccg gacatcagcg ctagcggagt gtatactggc ttactatgtt
      481 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
      541 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
      601 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
      661 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
      721 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
      781 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
      841 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
      901 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
      961 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1021 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     1081 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     1141 ccaagccatt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     1201 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     1261 gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca
     1321 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg tttgacagct
     1381 tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc
     1441 accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc
     1501 tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc
     1561 cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct
     1621 atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc
     1681 ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat
     1741 cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
     1801 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc
     1861 ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc
     1921 cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg
     1981 cttcctaatg caggagtcgc ataagggaga gcgtcgaccg atgcccttga gagccttcaa
     2041 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     2101 tttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     2161 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     2221 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     2281 ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac
     2341 gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc
     2401 cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg
     2461 atcgctcgcg gctcttacca gcctaacttc gatcattgga ccgctgatcg tcacggcgat
     2521 ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg ccgccctata
     2581 ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct cgacctgaat
     2641 ggaagccggc ggcacctcgc taacggattc accactccaa gaattggagc caatcaattc
     2701 ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga acatatccat cgcgtccgcc
     2761 atctccagca gccgcacgcg gcgcatctcg ggcagcgttg ggtcctggcc acgggtgcgc
     2821 atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag
     2881 cagaatgaat caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg
     2941 acctgagcgc cgcagccgga cgaccgagcg ccttggactc ctgttgatag atccagtaat
     3001 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     3061 tggtgagaat ggtctcaaat gcagagaaga gccagtaggg cagtgagcgc aacgcaatta
     3121 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     3181 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     3241 acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat
     3301 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3361 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3421 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     3481 cttacgca
//
