LOCUS       sequence-215498-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 1DE 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: pStA1DE
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..95
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    144..165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(180..185,186..203,204..210)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    218..234
                     /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     223..245
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     242..258
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     242..258
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             254..577
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    271..327
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(328..345)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(328..344)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(337..359)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(553..572)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(605..691)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1094..1111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1228..1773)
                     /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(1308..1327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1885..1890,1891..1907,1908..1913)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1961..3151
                     /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(2313..2332)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
       61 tcggttgccg ccgggcgttt tttattggtg agaatggtct cagcttcaga gaagagccag
      121 tagggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
      181 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
      241 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
      301 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
      361 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      421 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      481 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
      541 gtgcactctc agtacaatct gctctgatgc cgcatagttt ccgctcttca ggacgctaga
      601 gaccatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc
      661 agtctttcga ctgagccttt cgttttattt gatgcccgaa agggcctcgt gatacgccaa
      721 aagttggccc agggcttccc ggtatcaaca gggacaccag gatttattta ttctgcgaag
      781 tgatcttccg tcacaggtat ttattcggcg caaagtgcgt cgggtgatgc tgccaactta
      841 ctgatttagt gtatgatggt gtttttgagg tgctccagtg gcttctgttt ctatcagctg
      901 tccctcctgt tcagctactg acggggtggt gcgtaacggc aaaagcaccg ccggacatca
      961 gcgctagcgg agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg
     1021 cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga
     1081 tatattccgc ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg
     1141 aaatggctta cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg
     1201 aagtgagagg gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca
     1261 cgaaatctga cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc
     1321 gtttccccct ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt
     1381 cattccgctg ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg
     1441 cagttcgctc caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct
     1501 tatccggtaa ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag
     1561 cagccactgg taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa
     1621 actgaaagga caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag
     1681 ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag
     1741 caagagatta cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa
     1801 tatttctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     1861 cgatataagt tgtaattctc atgtttgaca gcttatcatc gataagcttt aatgcggtag
     1921 tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct
     1981 catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg ttatgccggt
     2041 actgccgggc ctcttgcggg atatcgtcca ttccgacagc atcgccagtc actatggcgt
     2101 gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca cccgttctcg gagcactgtc
     2161 cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta cttggagcca ctatcgacta
     2221 cgcgatcatg gcgaccacac ccgtcctgtg gatcctctac gccggacgca tcgtggccgg
     2281 catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga
     2341 agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg
     2401 ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc ttgcggcggc
     2461 ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt cgcataaggg
     2521 agagcgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     2581 gggcatgact atcgtcgccg cacttatgac tgttttcttt atcatgcaac tcgtaggaca
     2641 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     2701 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     2761 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga
     2821 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     2881 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     2941 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac
     3001 ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa
     3061 cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
     3121 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
     3181 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
     3241 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
     3301 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     3361 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     3421 acgtgaagcg actgctgctg caaaacgtct gcgacctgag cgccgcagcc gaacgaccga
     3481 gcgccttg
//
