LOCUS       Exported                6620 bp ds-DNA     circular SYN 19-JUN-2024
DEFINITION  Shoebridge_S6J_6_pCAG_GST-TEV-WIPI4     coverage: 2.74e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6620)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6620
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(163..179)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    187..203
                     /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)."
     promoter        complement(211..241)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    256..277
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        336..531
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      382..517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    537..671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(910..1498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1669..2529)
                     /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"
     promoter        complement(2530..2634)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2665..3044
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3046..3322
                     /label=chicken beta-actin promoter
     intron          3323..4339
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      4413..4422
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4419..5075
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             5103..5123
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5130..6212
                     /codon_start=1
                     /label=WIPI4
                     /translation="MTQQPLRGVTSLRFNQDQSCFCCAMETGVRIYNVEPLMEKGHLDH
                     EQVGSMGLVEMLHRSNLLALVGGGSSPKFSEISVLIWDDAREGKDSKEKLVLEFTFTKP
                     VLSVRMRHDKIVIVLKNRIYVYSFPDNPRKLFEFDTRDNPKGLCDLCPSLEKQLLVFPG
                     HKCGSLQLVDLASTKPGTSSAPFTINAHQSDIACVSLNQPGTVVASASQKGTLIRLFDT
                     QSKEKLVELRRGTDPATLYCINFSHDSSFLCASSDKGTVHIFALKDTRLNRRSALARVG
                     KVGPMIGQYVDSQWSLASFTVPAESACICAFGRNTSKNVNSVIAICVDGTFHKYVFTPD
                     GNCNREAFDVYLDICDDDDF"
     polyA_signal    6367..6422
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
ORIGIN
        1 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
       61 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
      121 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
      181 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
      241 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
      301 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
      361 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      421 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      481 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
      541 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
      601 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
      661 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      721 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
      781 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
      841 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
      901 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
      961 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1021 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1081 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1141 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1201 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1261 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1321 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1381 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1441 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1501 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1561 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1621 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1681 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1741 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1801 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1861 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1921 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1981 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2041 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2101 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     2161 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     2221 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     2281 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     2341 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     2401 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     2461 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     2521 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2581 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2641 tccccgaaaa gtgccacctg ggtcgacatt gattattgac tagttattaa tagtaatcaa
     2701 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     2761 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2821 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2881 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2941 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3001 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     3061 cgttctgctt cactctcccc atctcccccc ctccccaccc ccaattttgt atttatttat
     3121 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
     3181 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     3241 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     3301 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     3361 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     3421 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     3481 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     3541 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     3601 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     3661 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     3721 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     3781 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     3841 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     3901 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     3961 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     4021 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     4081 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     4141 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     4201 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     4261 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     4321 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     4381 tggcaaagaa ttcctcgatc gacggtatcg atgccaccat ggcatcccct atactaggtt
     4441 attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat cttgaagaaa
     4501 aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac aaaaagtttg
     4561 aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt aaattaacac
     4621 agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt ggttgtccaa
     4681 aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga tacggtgttt
     4741 cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt agcaagctac
     4801 ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta aatggtgatc
     4861 atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta tacatggacc
     4921 caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt gaagctatcc
     4981 cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg cagggctggc
     5041 aagccacgtt tggtggtggc gaccatcctc caaaatcgga tgattacgat atcccaacga
     5101 ccgaaaacct gtattttcag ggtggtacca tgactcaaca gccacttcga ggagtgacca
     5161 gcctgcgttt caaccaagac caaagctgct tttgctgcgc catggagaca ggtgtgcgca
     5221 tctacaacgt ggagcccttg atggagaagg ggcatctgga ccacgagcag gtgggcagca
     5281 tgggcttggt ggagatgctg caccgctcca accttctggc cttggtgggc ggtggtagta
     5341 gtcccaagtt ctcagagatc tcagtgctga tctgggacga tgcccgggag ggcaaggact
     5401 ccaaggagaa gctggtgctg gagttcacct tcaccaagcc agtgctttct gtgcgcatgc
     5461 gccatgacaa gatcgtgatc gtgctgaaga accgcatcta tgtgtactcc ttccccgaca
     5521 atccccgaaa gctgtttgag tttgataccc gggacaaccc caaggggctc tgtgacctct
     5581 gccccagcct ggagaagcaa ctgctagtgt tcccgggaca caagtgtggg agtctgcaac
     5641 ttgtggacct ggcgagcaca aagcctggca cctcgtctgc tccattcacg atcaatgcac
     5701 atcagagtga catagcctgt gtgtctctaa accagccagg cactgtagtg gcctcagcct
     5761 cccagaaggg tacccttatt cgcctctttg acacacaatc caaggagaaa ctggtggagc
     5821 tgcgccgagg cactgaccct gccaccctct actgcattaa cttcagccac gactcctcct
     5881 tcctctgcgc ttccagtgat aagggtactg tccatatctt tgctctcaag gatacccgcc
     5941 tcaaccgccg ctccgcgctg gctcgcgtgg gcaaggtggg gcctatgatt gggcagtacg
     6001 tggactctca gtggagcctg gcgagcttca ctgtgcctgc tgagtcagct tgcatctgcg
     6061 ccttcggtcg caatacttcc aagaacgtca actctgtcat tgccatctgc gtagatggga
     6121 ccttccacaa atatgtcttc actcctgatg gaaactgcaa cagagaggct ttcgacgtgt
     6181 accttgacat ctgtgatgat gatgactttt aactcgagga attcactcct caggtgcagg
     6241 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     6301 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     6361 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     6421 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     6481 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     6541 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     6601 cttgaggtta gatttttttt
//
