LOCUS       sequence-229109-        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"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /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     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             75..398
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    92..148
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(149..166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(149..165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(158..180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(374..393)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(426..512)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(915..932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1049..1594)
                     /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(1129..1148)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1706..1711,1712..1728,1729..1734)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1782..2972
                     /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(2134..2153)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      3310..3404
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    3453..3474
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
      121 aggatccccg ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
      181 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      241 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      301 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      361 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt tccgctcttc aggacgctag
      421 agaccatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
      481 cagtctttcg actgagcctt tcgttttatt tgatgcccga aagggcctcg tgatacgcca
      541 aaagttggcc cagggcttcc cggtatcaac agggacacca ggatttattt attctgcgaa
      601 gtgatcttcc gtcacaggta tttattcggc gcaaagtgtg tcgggtgatg ctgccaactt
      661 actgatttag tgtatgatgg tgtttttgag gtgctccagt ggcttctgtt tctatcagct
      721 gtccctcctg ttcagctact gacggggtgg tgcgtaacgg caaaagcacc gccggacatc
      781 agcgctagcg gagtgtatac tggcttacta tgttggcact gatgagggtg tcagtgaagt
      841 gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc agcagaatat gtgatacagg
      901 atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact gcgacgagcg
      961 gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat acttaacagg
     1021 gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct gacaagcatc
     1081 acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa agataccagg
     1141 cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg
     1201 tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag
     1261 gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc
     1321 ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca
     1381 gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta
     1441 aactgaaagg acaagttttg gtgactgcgc tcctccaagc catttacctc ggttcaaaga
     1501 gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga
     1561 gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa
     1621 atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat
     1681 acgatataag ttgtaattct catgtttgac agcttatcat cgataagctt taatgcggta
     1741 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     1801 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     1861 tactgccggg cctcttgcgg gatatcgtcc attccgacag catcgccagt cactatggcg
     1921 tgctgctagc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc ggagcactgt
     1981 ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc actatcgact
     2041 acgcgatcat ggcgaccaca cccgtcctgt ggatcctcta cgccggacgc atcgtggccg
     2101 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
     2161 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
     2221 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
     2281 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
     2341 gagagcgtcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
     2401 ggggcatgac tatcgtcgcc gcacttatga ctgttttctt tatcatgcaa ctcgtaggac
     2461 aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga
     2521 tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca
     2581 ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc atggcggccg
     2641 acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc ttccccatta
     2701 tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc
     2761 aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt accagcctaa
     2821 cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg agcacatgga
     2881 acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc
     2941 gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg
     3001 attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga atgcgcaaac
     3061 caacccttgg cagaacatat ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat
     3121 ctcgggcagc gttgggtcct ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac
     3181 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
     3241 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcgccgcagc cggacgaccg
     3301 agcgccttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     3361 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatggtctc agcttcagag
     3421 aagagccagt agggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     3481 ccccaggc
//
