LOCUS       sequence-229065-        3040 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 212 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3040)
  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 3040)
  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: pStA212
FEATURES             Location/Qualifiers
     source          1..3040
                     /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(463..549)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      899..1444
                     /direction=RIGHT
                     /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     1345..1364
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1561..1578
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2028..2047)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2039..2854
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2039..2854
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2112..2131)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2897..2991
                     /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 gtctcatact actaaagtct tcatttgtcc tactcaggag
      481 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
      541 ttttatttga tgcccgaaag ggcctcgtga tacgcaaatc tggagccggt gagcgcggta
      601 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
      661 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
      721 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
      781 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
      841 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cttaataaga tgatcttctt
      901 gagatcgttt tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg
      961 gcggtttttc gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag
     1021 gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga
     1081 ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc
     1141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt
     1201 tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga
     1261 atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg
     1321 agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1381 tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg
     1441 gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc
     1501 aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt
     1561 agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc
     1621 ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa
     1681 catagtaagc cagtatacac tccgctagcg ctgaggtctg cctcgtgaag aaggtgttgc
     1741 tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt
     1801 gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga
     1861 acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt
     1921 tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     1981 tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2041 gagccatatt caacgggaaa cgtcttgctc gaggccgcga ttaaattcca acatggatgc
     2101 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2161 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2221 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2281 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2341 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2401 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2461 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2521 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2581 aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     2641 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     2701 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     2761 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     2821 gcagtttcat ttgatgctcg atgagttttt ctaatcttgg gggaaattag ccgcagccga
     2881 acgaccgagc gccttggact cctgttgata gatccagtaa tgacctcaga actccatctg
     2941 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tgaagacaat
     3001 gccggagaga gacccagtag ggcagtgagc gcaacgcaat
//
