LOCUS       sequence-229069-        3197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Start-Stop Assembly Level 313 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: pStA313
FEATURES             Location/Qualifiers
     source          1..3197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..18)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1..17)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10..32)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(226..245)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      complement(268..354)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(856..1515)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1430..1449
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1516..1618)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2010..2027)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2144..2689)
                     /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(2224..2243)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2801..2806,2807..2823,2824..2829)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      2881..2975
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    3014..3035
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3050..3055,3056..3073,3074..3080)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3088..3104
                     /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     3093..3115
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3112..3128
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3112..3128
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(3124..3197,1..250)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    3141..3197
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
       61 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
      121 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
      181 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
      241 tgccgcatag tttccgaaga caattacatt tgtcctactc aggagagcgt tcaccgacaa
      301 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgccc
      361 gaaagggcct cgtgatacgc ctttatcatg caactcgtag gacaggtgcc ggcagcgccc
      421 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gccctgcacc
      481 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
      541 gtattaacga agcgctaacc gtttttatca ggctctggga ggcagaataa atgatcatat
      601 cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca tttgagaagc
      661 acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg
      721 ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc
      781 attcatccgc ttattatcac ttattcaggc gtagcaacca ggcgtttaag ggcaccaata
      841 actgccttaa aaaaattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt
      901 aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg
      961 catcagcacc ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa
     1021 gttgtccata ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga
     1081 gacgaaaaac atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca
     1141 cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca
     1201 gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc
     1261 ccatatcacc agctcaccgt ctttcattgc catacggaat tccggatgag cattcatcag
     1321 gcgggcaaga atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt
     1381 taaaaaggcc gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg
     1441 aaatgcctca aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt
     1501 gatttttttc tccattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     1561 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaac
     1621 gtctcatttt cgccaaaagt tggcccaggg cttcccggta tcaacaggga caccaggatt
     1681 tatttattct gcgaagtgat cttccgtcac aggtatttat tcggcgcaaa gtgcgtcggg
     1741 tgatgctgcc aacttactga tttagtgtat gatggtgttt ttgaggtgct ccagtggctt
     1801 ctgtttctat cagctgtccc tcctgttcag ctactgacgg ggtggtgcgt aacggcaaaa
     1861 gcaccgccgg acatcagcgc tagcggagtg tatactggct tactatgttg gcactgatga
     1921 gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg ctgcaccggt gcgtcagcag
     1981 aatatgtgat acaggatata ttccgcttcc tcgctcactg actcgctacg ctcggtcgtt
     2041 cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga agatgccagg
     2101 aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat aggctccgcc
     2161 cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac ccgacaggac
     2221 tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct gttcctgcct
     2281 ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc acgcctgaca
     2341 ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc ccccgttcag
     2401 tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga aagacatgca
     2461 aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct tgaagtcatg
     2521 cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc caagccagtt
     2581 acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct gcaaggcggt
     2641 tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag aagatcatct
     2701 tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa atgtagcacc
     2761 tgaagtcagc cccatacgat ataagttgta attctcatgt ttgacagctt atcatcgata
     2821 agctttaatg cggtagttta tcacagttaa attgccgcag ccgaacgacc gagcgccttg
     2881 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     2941 tcggttgccg ccgggcgttt tttattggtg agaattgcca agtcttccag tagggcagtg
     3001 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3061 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     3121 gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga ggatccccgg
     3181 gtaccgagct cgaattc
//
