LOCUS       Exported                6570 bp ds-DNA     circular SYN 19-JUN-2024
DEFINITION  Shoebridge_S6J_5_pCAG_GST-TEV-WIPI3     coverage: 2.89e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6570)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6570
                     /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..3320
                     /label=chicken beta-actin promoter
     intron          3321..4337
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      4411..4420
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4417..5073
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             5101..5121
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5128..6162
                     /codon_start=1
                     /label=WIPI3
                     /translation="MNLLPCNPHGNGLLYAGFNQDHGCFACGMENGFRVYNTDPLKEKE
                     KQEFLEGGVGHVEMLFRCNYLALVGGGKKPKYPPNKVMIWDDLKKKTVIEIEFSTEVKA
                     VKLRRDRIVVVLDSMIKVFTFTHNPHQLHVFETCYNPKGLCVLCPNSNNSLLAFPGTHT
                     GHVQLVDLASTEKPPVDIPAHEGVLSCIALNLQGTRIATASEKGTLIRIFDTSSGHLIQ
                     ELRRGSQAANIYCINFNQDASLICVSSDHGTVHIFAAEDPKRNKQSSLASASFLPKYFS
                     SKWSFSKFQVPSGSPCICAFGTEPNAVIAICADGSYYKFLFNPKGECIRDVYAQFLEMT
                     DDKL"
     polyA_signal    6317..6372
                     /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 ggggcgcgcg ccaggcgggg
     3181 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     3241 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     3301 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
     3361 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     3421 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     3481 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     3541 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
     3601 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
     3661 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     3721 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     3781 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     3841 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     3901 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     3961 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     4021 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     4081 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     4141 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     4201 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     4261 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
     4321 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
     4381 gcaaagaatt cctcgatcga cggtatcgat gccaccatgg catcccctat actaggttat
     4441 tggaaaatta agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa
     4501 tatgaagagc atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa
     4561 ttgggtttgg agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag
     4621 tctatggcca tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa
     4681 gagcgtgcag agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg
     4741 agaattgcat atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct
     4801 gaaatgctga aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat
     4861 gtaacccatc ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca
     4921 atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca
     4981 caaattgata agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa
     5041 gccacgtttg gtggtggcga ccatcctcca aaatcggatg attacgatat cccaacgacc
     5101 gaaaacctgt attttcaggg tggtaccatg aatctgctgc cttgtaatcc acacgggaac
     5161 gggctgctgt atgctgggtt taatcaggac cacggctgct tcgcttgcgg aatggagaac
     5221 gggttcagag tgtacaatac cgacccactg aaagagaagg aaaaacagga gttcctggaa
     5281 ggcggggtcg gacatgtgga aatgctgttt aggtgcaact acctggctct ggtgggaggc
     5341 gggaagaaac ccaagtatcc ccctaacaaa gtgatgatct gggacgatct gaagaaaaag
     5401 actgtgatcg agattgagtt cagcaccgag gtcaaggccg tgaaactgcg gagagaccgc
     5461 atcgtggtcg tgctggattc catgatcaag gtcttcacct ttacacacaa ccctcatcag
     5521 ctgcacgtgt tcgaaacatg ctataacccc aagggcctgt gcgtgctgtg ccctaattca
     5581 aacaatagcc tgctggcttt tcccggaact cataccggcc acgtccagct ggtggacctg
     5641 gcatctactg agaagccacc cgtcgatatc cctgcacatg aaggggtgct gtcctgtatc
     5701 gccctgaatc tgcagggaac acgaattgct actgcatctg agaagggcac cctgatccgg
     5761 attttcgaca caagctccgg ccacctgatt caggaactga ggcgcgggag tcaggccgct
     5821 aacatctact gcattaactt caatcaggac gccagcctga tctgcgtgag cagtgatcat
     5881 ggcaccgtgc acatttttgc agccgaggat ccaaagcgaa acaagcagtc aagcctggcc
     5941 agtgcttcat tcctgcccaa gtatttttcc tctaaatgga gcttctccaa gtttcaggtg
     6001 ccttctggga gtccatgcat ctgtgccttt ggaacagagc ctaacgcagt gatcgccatt
     6061 tgtgctgatg gcagctacta taaattcctg tttaatccaa agggggagtg tattagggat
     6121 gtctatgctc agttcctgga gatgaccgac gataagctgt aactcgagga attcactcct
     6181 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     6241 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     6301 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     6361 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     6421 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     6481 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     6541 aaagccttga cttgaggtta gatttttttt
//
