LOCUS       sequence-229093-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 1CD 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: pStA1CD
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..18)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1..17)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10..32)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(226..245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(278..364)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(767..784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(901..1446)
                     /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(981..1000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1558..1563,1564..1580,1581..1586)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1634..2824
                     /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(1986..2005)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      3162..3256
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    3305..3326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3341..3346,3347..3364,3365..3371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3379..3395
                     /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     3384..3406
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3403..3419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3403..3419
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3415..3488,1..250)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    3432..3488
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
       61 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      121 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      181 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
      241 tgccgcatag tttccgctct tcaggagctt agagaccatt tgtcctactc aggagagcgt
      301 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
      361 tttgatgccc gaaagggcct cgtgatacgc caaaagttgg cccagggctt cccggtatca
      421 acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg tatttattcg
      481 gcgcaaagtg tgtcgggtga tgctgccaac ttactgattt agtgtatgat ggtgtttttg
      541 aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta ctgacggggt
      601 ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat actggcttac
      661 tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga aaaaaggctg
      721 caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg ctcactgact
      781 cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg ggcggagatt
      841 tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt
      901 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
      961 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
     1021 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
     1081 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
     1141 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1201 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
     1261 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
     1321 gctcctccaa gccatttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
     1381 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
     1441 tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt gcaatttatc
     1501 tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt ctcatgtttg
     1561 acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag
     1621 tcaggcaccg tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct
     1681 ggatgctgta ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatcgt
     1741 ccattccgac agcatcgcca gtcactatgg cgtgctgcta gcgctatatg cgttgatgca
     1801 atttctatgc gcacccgttc tcggagcact gtccgaccgc tttggccgcc gcccagtcct
     1861 gctcgcttcg ctacttggag ccactatcga ctacgcgatc atggcgacca cacccgtcct
     1921 gtggatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
     1981 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
     2041 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
     2101 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
     2161 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgaccgatgc ccttgagagc
     2221 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2281 gactgttttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     2341 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     2401 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     2461 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     2521 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     2581 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     2641 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac
     2701 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     2761 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     2821 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     2881 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     2941 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg
     3001 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     3061 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     3121 tctgcgacct gagcgccgca gccggacgac cgagcgcctt ggactcctgt tgatagatcc
     3181 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     3241 ttttattggt gagaatggtc tcaaggtcag agaagagcca gtagggcagt gagcgcaacg
     3301 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     3361 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     3421 atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
     3481 tcgaattc
//
