LOCUS       Exported                6565 bp ds-DNA     circular SYN 19-JUN-2024
DEFINITION  Shoebridge_S6J_1_pCAG_GST-TEV-WIPI1_peak_2     coverage: 2.84e+03x.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6565
                     /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."
     rep_origin      complement(577..1165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1336..2196)
                     /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(2197..2301)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2332..2711
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2713..2988
                     /label=chicken beta-actin promoter
     intron          2989..4005
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     regulatory      4081..4090
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4087..4743
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             4771..4791
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4804..6144
                     /codon_start=1
                     /label=WIPI1
                     /translation="MEAEAADAPPGGVESALSCFSFNQDCTSLATGTKAGYKLFSLSSV
                     EQLDQVHGSNEIPDVYIVERLFSSSLVVVVSHTKPRQMNVYHFKKGTEICNYSYSSNIL
                     SIRLNRQRLLVCLEESIYIHNIKDMKLLKTLLDIPANPTGLCALSINHSNSYLAYPGSL
                     TSGEIVLYDGNSLKTVCTIAAHEGTLAAITFNASGSKLASASEKGTVIRVFSVPDGQKL
                     YEFRRGMKRYVTISSLVFSMDSQFLCASSNTETVHIFKLEQVTNSRPEEPSTWSGYMGK
                     MFMAATNYLPTQVSDMMHQDRAFATARLNFSGQRNICTLSTIQKLPRLLVASSSGHLYM
                     YNLDPQDGGECVLIKTHSLLGSGTTEENKENDLRPSLPQSYAATVARPSASSASTVPGY
                     SEDGGALRGEVIPEHEFATGPVCLDDENEFPPIILCRGNQKGKTKQS"
     polyA_signal    6312..6367
                     /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 gatcgatccg ctgcattaat gaatcggcca
      361 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
      421 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      481 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
      541 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
      601 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
      661 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
      721 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
      781 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
      841 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
      901 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
      961 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1021 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1081 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1141 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1201 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1261 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1321 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1381 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1441 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1501 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1561 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1621 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1681 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1741 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1801 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     1861 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     1921 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     1981 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2041 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2101 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2161 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2221 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2281 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt cgacattgat
     2341 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     2401 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     2461 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     2521 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     2581 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     2641 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     2701 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tccccccctc
     2761 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     2821 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     2881 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     2941 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     3001 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
     3061 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
     3121 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     3181 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     3241 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     3301 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     3361 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     3421 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     3481 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     3541 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     3601 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     3661 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     3721 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     3781 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     3841 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     3901 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     3961 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     4021 ctggttattg tgctgtctca tcattttggc aaagaattgc ggccgtctca ggccgaattc
     4081 gccaccatgg catcccctat actaggttat tggaaaatta agggccttgt gcaacccact
     4141 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
     4201 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
     4261 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
     4321 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
     4381 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
     4441 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
     4501 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
     4561 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
     4621 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
     4681 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
     4741 aaatcggatg attacgatat cccaacgacc gaaaacctgt attttcaggg tgtcgacggt
     4801 accatggaag ccgaagccgc tgatgctcct cctggcggag ttgaatctgc cctgagctgc
     4861 ttcagcttca accaggactg taccagcctg gccacaggca caaaggccgg ctacaagctg
     4921 tttagcctga gcagcgtgga acagctggat caggtgcacg gcagcaacga gatccccgac
     4981 gtgtacatcg tggaacggct gttcagcagc agcctggtgg tggtcgtgtc tcacaccaag
     5041 cctcggcaga tgaacgtgta ccacttcaag aagggcaccg aaatctgcaa ctacagctac
     5101 agcagcaaca tcctgagcat ccggctgaac cggcagagac tgctcgtgtg tctggaagag
     5161 agcatctaca tccacaacat caaggacatg aagctgctca agaccctgct ggacatcccc
     5221 gccaatccta caggactgtg tgccctgtcc atcaaccaca gcaacagcta cctggcttac
     5281 cccggctctc tgacatctgg cgagatcgtg ctgtacgacg gcaacagcct gaaaaccgtg
     5341 tgcacaatcg ccgctcacga gggaacactg gccgccatca cctttaatgc cagcggctct
     5401 aaactggcca gcgccagcga aaagggcaca gtgatcagag tgttcagcgt gcccgatggc
     5461 cagaagctgt acgagtttcg gagaggcatg aagcgctacg tgaccatcag ctccctggtg
     5521 ttctctatgg acagccagtt cctgtgcgcc agctccaaca cagagacagt gcacatcttc
     5581 aagctggaac aagtgaccaa cagcagaccc gaggaaccct ctacttggag cggctacatg
     5641 ggcaagatgt tcatggccgc caccaactac ctgcctacac aggtgtccga catgatgcac
     5701 caggaccggg cctttgccac cgccagactg aatttttccg gccagcggaa catctgcacc
     5761 ctgagcacca ttcagaagct gcccagactg ctggtggcct cttctagcgg ccacctgtac
     5821 atgtataatc tggaccctca ggacggcggc gagtgcgtgc tgattaagac acactctctg
     5881 ctcggcagcg gcaccaccga ggaaaacaaa gagaacgacc tgcggcctag cctgcctcag
     5941 tcttatgctg ccacagtggc cagacctagc gcctcttctg cttctacagt gcccggctat
     6001 agcgaggatg gcggagcact tagaggcgaa gtgatccccg agcacgagtt cgctacaggc
     6061 cctgtctgtc tggacgacga gaacgagttc cctcctatca tcctgtgccg gggcaaccag
     6121 aagggcaaga ccaagcagtc ttgactcgag actagtatcg cgataattca ctcctcaggt
     6181 gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca
     6241 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     6301 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     6361 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     6421 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     6481 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     6541 cttgacttga ggttagattt ttttt
//
