LOCUS       Exported                2658 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Start-Stop Assembly Level 0 plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    pStA0
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2658
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     395..414
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      652..746
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     protein_bind    785..806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        821..851
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    859..875
                     /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     864..886
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     883..899
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     883..899
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             895..1218
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    912..968
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(969..986)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(969..985)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(978..1000)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1194..1213)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      1243..1329
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1342..1360)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1428..1532
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1533..2393
                     /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(1751..1770)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2564..494
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
       61 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      121 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      181 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      241 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      301 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      361 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      421 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      481 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      541 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      601 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcctt ggactcctgt
      661 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
      721 gccgggcgtt ttttattggt gagaatccag tgtgagagac caagggcagt gagcgcaacg
      781 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
      841 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      901 atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
      961 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     1021 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     1081 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     1141 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     1201 tgctctgatg ccgcataggg tctcagacct ctagggcggc ggatttgtcc tactcaggag
     1261 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     1321 ttttatttga tgcccgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     1381 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1441 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1501 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1561 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1621 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1681 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1741 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1801 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1861 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1921 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1981 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2041 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2101 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2161 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2221 ctgataaatc tggagccggt gagcgtgggt cccgcggtat cattgcagca ctggggccag
     2281 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2341 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2401 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2461 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2641 cggatcaaga gctaccaa
//
