LOCUS       sequence-237139-        3488 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 1DZ 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: pStA1DZ
FEATURES             Location/Qualifiers
     source          1..3488
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..324
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    18..74
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(75..92)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(75..91)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(84..106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(300..319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(352..438)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(841..858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(975..1520)
                     /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(1055..1074)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1632..1637,1638..1654,1655..1660)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1708..2898
                     /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(2060..2079)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     terminator      3236..3330
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    3379..3400
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3415..3420,3421..3438,3439..3445)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3453..3469
                     /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     3458..3480
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3477..3488,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3477..3488,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      301 acaatctgct ctgatgccgc atagtttccg ctcttcagga tactagagac catttgtcct
      361 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
      421 agcctttcgt tttatttgat gcccgaaagg gcctcgtgat acgccaaaag ttggcccagg
      481 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
      541 caggtattta ttcggcgcaa agtgtgtcgg gtgatgctgc caacttactg atttagtgta
      601 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
      661 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
      721 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
      781 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
      841 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
      901 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
      961 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     1021 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
     1081 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
     1141 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
     1201 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
     1261 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
     1321 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
     1381 gttttggtga ctgcgctcct ccaagccatt tacctcggtt caaagagttg gtagctcaga
     1441 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
     1501 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt
     1561 cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt
     1621 aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta
     1681 aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc
     1741 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
     1801 ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta
     1861 tatgcgytga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc
     1921 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg
     1981 accacacccg tcctgtggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
     2041 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
     2101 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
     2161 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
     2221 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaccg
     2281 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     2341 gtcgccgcac ttatgactgt tttctttatc atgcaactcg taggacaggt gccggcagcg
     2401 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     2461 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     2521 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
     2581 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
     2641 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
     2701 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcattgga
     2761 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
     2821 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
     2881 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
     2941 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
     3001 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg
     3061 ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
     3121 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
     3181 gctgctgcaa aacgtctgcg acctgagcgc cgcagccgga cgaccgagcg ccttggactc
     3241 ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt
     3301 tgccgccggg cgttttttat tggtgagaat ggtctcagct tcagagaaga gccagtaggg
     3361 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     3421 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     3481 aaacagct
//
