LOCUS       sequence-215490-        2658 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 0 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2658)
  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 2658)
  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: pStA0
FEATURES             Location/Qualifiers
     source          1..2658
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1..95
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    134..155
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(170..175,176..193,194..200)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    208..224
                     /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     213..235
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     232..248
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     232..248
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             244..567
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    261..317
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(318..335)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(318..334)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(327..349)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(543..562)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(592..678)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(691..709)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        777..881
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(882..950,951..1742)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1100..1119)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1913..2501
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2402..2421
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
       61 tcggttgccg ccgggcgttt tttattggtg agaatccagt gtgagagacc aagggcagtg
      121 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
      181 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
      241 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
      301 gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
      361 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
      421 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
      481 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
      541 agtacaatct gctctgatgc cgcatagggt ctcagacctc tagggcggcg gatttgtcct
      601 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
      661 agcctttcgt tttatttgat gcccgaaagg gcctcgtgat acgcctattt ttataggtta
      721 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
      781 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      841 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      901 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      961 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1021 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1081 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1141 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     1201 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     1261 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1321 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1381 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1441 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1501 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1561 ggtttattgc tgataaatct ggagccggtg agcgtgggtc ccgcggtatc attgcagcac
     1621 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1681 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1741 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1801 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1861 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1921 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1981 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2041 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2101 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2161 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2221 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2281 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2341 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2401 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2461 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2521 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2581 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2641 gaacgaccga gcgccttg
//
