LOCUS       sequence-229092-        3197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 314 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3197)
  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 3197)
  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: pStA314
FEATURES             Location/Qualifiers
     source          1..3197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             111..434
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    128..184
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(185..202)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(185..201)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(194..216)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(410..429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(452..538)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1040..1699)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1614..1633
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1700..1802)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2194..2211)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2328..2873)
                     /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(2408..2427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2985..2990,2991..3007,3008..3013)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      3065..3159
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      121 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
      181 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      241 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      301 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
      361 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      421 ctgatgccgc atagtttccg aagacaacga gatttgtcct actcaggaga gcgttcaccg
      481 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
      541 gcccgaaagg gcctcgtgat acgcctttat catgcaactc gtaggacagg tgccggcagc
      601 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgccctg
      661 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac
      721 atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga ataaatgatc
      781 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag
      841 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa
      901 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc
      961 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc
     1021 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     1081 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1141 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     1201 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     1261 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     1321 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     1381 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     1441 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     1501 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     1561 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     1621 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     1681 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1741 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     1801 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag
     1861 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt
     1921 cgggtgatgc tgccaactta ctgatttagt gtatgatggt gtttttgagg tgctccagtg
     1981 gcttctgttt ctatcagctg tccctcctgt tcagctactg acggggtggt gcgtaacggc
     2041 aaaagcaccg ccggacatca gcgctagcgg agtgtatact ggcttactat gttggcactg
     2101 atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac cggtgcgtca
     2161 gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc tacgctcggt
     2221 cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc tggaagatgc
     2281 caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt ccataggctc
     2341 cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg aaacccgaca
     2401 ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc tcctgttcct
     2461 gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca ttccacgcct
     2521 gacactcagt ttcgggtagg cagttcgctc caagctggac tgtatgcacg aaccccccgt
     2581 tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggaaagaca
     2641 tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta gtcttgaagt
     2701 catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct cctccaagcc
     2761 agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg ccctgcaagg
     2821 cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct caagaagatc
     2881 atcttattaa tcagataaaa tatttctaga tttcagtgca atttatctct tcaaatgtag
     2941 cacctgaagt cagccccata cgatataagt tgtaattctc atgtttgaca gcttatcatc
     3001 gataagcttt aatgcggtag tttatcacag ttaaattgcc gcagccgaac gaccgagcgc
     3061 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     3121 acgctcggtt gccgccgggc gttttttatt ggtgagaatt gccaagtctt ccagtagggc
     3181 agtgagcgca acgcaat
//
