LOCUS       Exported                3196 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Start-Stop Assembly Level 313 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pStA313
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3196)
  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 3196)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3196
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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        170..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      585..671
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1172..1831)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1746..1765
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1832..1934)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(2326..2343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2460..3005)
                     /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(2540..2559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3117..3145
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
       61 tcggttgccg ccgggcgttt tttattggtg agaattgcca agtcttccag tagggcagtg
      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 cgcatagttt ccgaagacaa ttacatttgt cctactcagg
      601 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
      661 cgttttattt gatgcccgaa agggcctcgt gatacgcctt tatcatgcaa ctcgtaggac
      721 aggtgccggc agcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
      781 aacaagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
      841 gtggaacacc tacatctgta ttaacgaagc gctaaccgtt tttatcaggc tctgggaggc
      901 agaataaatg atcatatcgt caattattac ctccacgggg agagcctgag caaactggcc
      961 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     1021 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     1081 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaccaggcg
     1141 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
     1201 actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg gcatgatgaa
     1261 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga
     1321 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     1381 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     1441 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     1501 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     1561 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cggaattccg
     1621 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     1681 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     1741 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     1801 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     1861 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     1921 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
     1981 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg
     2041 cgcaaagtgc gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
     2101 ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg
     2161 gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact
     2221 atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc
     2281 accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc
     2341 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     2401 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     2461 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     2581 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     2641 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     2701 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2761 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     2821 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     2881 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     2941 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     3001 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     3061 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc tcatgtttga
     3121 cagcttatca tcgataagct ttaatgcggt agtttatcac agttaaattg ccgcagccga
     3181 acgaccgagc gccttg
//
