LOCUS       sequence-237130-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 1BZ 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: pStA1BZ
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(330..347)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(464..1009)
                     /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(544..563)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1121..1126,1127..1143,1144..1149)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1197..2387
                     /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(1549..1568)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      2725..2819
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    2868..2889
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2904..2909,2910..2927,2928..2934)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2942..2958
                     /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     2947..2969
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2966..2982
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2966..2982
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2978..3301
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2995..3051
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3052..3069)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3052..3068)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3061..3083)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3277..3296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(3329..3415)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgtgtcggg
       61 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
      121 ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa
      181 gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
      241 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
      301 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
      361 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
      421 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
      481 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
      541 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
      601 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
      661 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
      721 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
      781 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
      841 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccattt
      901 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
      961 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     1021 tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc
     1081 tgaagtcagc cccatacgat ataagttgta attctcatgt ttgacagctt atcatcgata
     1141 agctttaatg cggtagttta tcacagttaa attgctaacg cagtcaggca ccgtgtatga
     1201 aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac cctggatgct gtaggcatag
     1261 gcttggttat gccggtactg ccgggcctct tgcgggatat cgtccattcc gacagcatcg
     1321 ccagtcacta tggcgtgctg ctagcgctat atgcgttgat gcaatttcta tgcgcacccg
     1381 ttctcggagc actgtccgac cgctttggcc gccgcccagt cctgctcgct tcgctacttg
     1441 gagccactat cgactacgcg atcatggcga ccacacccgt cctgtggatc ctctacgccg
     1501 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     1561 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     1621 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
     1681 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
     1741 aggagtcgca taagggagag cgtcgaccga tgcccttgag agccttcaac ccagtcagct
     1801 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtt ttctttatca
     1861 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     1921 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     1981 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     2041 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     2101 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     2161 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg
     2221 ctcttaccag cctaacttcg atcattggac cgctgatcgt cacggcgatt tatgccgcct
     2281 cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc
     2341 tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     2401 gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
     2461 tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
     2521 ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
     2581 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     2641 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcgcc
     2701 gcagccggac gaccgagcgc cttggactcc tgttgataga tccagtaatg acctcagaac
     2761 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatg
     2821 gtctcaaatg cagagaagag ccagtagggc agtgagcgca acgcaattaa tgtgagttag
     2881 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     2941 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
     3001 gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg agctcgaatt cactggccgt
     3061 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     3121 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     3181 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     3241 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3301 gtttccgctc ttcaggatac tagagaccat ttgtcctact caggagagcg ttcaccgaca
     3361 aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt atttgatgcc
     3421 cgaaagggcc tcgtgatacg ccaaaagttg gcccagggct tcccggtatc aacagggaca
     3481 ccaggatt
//
