LOCUS       Exported                5845 bp ds-DNA     circular SYN 03-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5845)
  AUTHORS   eliasa55
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5845
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     149..166
                     /label=SMP_2198
     primer_bind     177..196
                     /label=SMP_1565
     primer_bind     179..201
                     /label=SMP_357
     promoter        183..211
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     primer_bind     183..202
                     /label=SMP_3645
     primer_bind     187..204
                     /label=SMP_2193
     protein_bind    219..235
                     /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     complement(227..266)
                     /label=Vector.REV
     primer_bind     238..252
                     /label=SMP_993
     primer_bind     249..285
                     /label=Fragment.FOR
     CDS             258..914
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MASPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
                     KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIR
                     YGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVV
                     LYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     258..276
                     /label=SMP2582
     regulatory      258..261
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(304..321)
                     /label=SMP2192
     primer_bind     complement(560..583)
                     /label=SMP1669
     primer_bind     799..820
                     /label=SMP358
     primer_bind     844..863
                     /label=GEX5-for
     primer_bind     875..892
                     /label=pETG-for
     primer_bind     876..895
                     /label=SMP3299
     primer_bind     complement(894..920)
                     /label=SMP2863
     primer_bind     925..941
                     /label=SMP2555
     CDS             942..962
                     /codon_start=1
                     /label=TEV cleavage site
                     /note="TEV cleavage site"
                     /translation="ENLYFQG"
     CDS             969..1685
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     969..988
                     /label=SMP2235
     primer_bind     969..988
                     /label=SMP3364
     regulatory      969..972
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     972..989
                     /label=SMP2824
     primer_bind     complement(972..989)
                     /label=SMP3729
     primer_bind     975..992
                     /label=SMP2016
     primer_bind     complement(1011..1027)
                     /label=PEN1-737R
     primer_bind     complement(1014..1035)
                     /label=SMP3301
     primer_bind     complement(1016..1036)
                     /label=SMP919
     primer_bind     complement(1031..1049)
                     /label=SMP2564
     primer_bind     complement(1042..1062)
                     /label=SMP2562
     primer_bind     complement(1044..1062)
                     /label=EGFP-N-R+33
     primer_bind     complement(1077..1095)
                     /label=EGFP-N-rev
     primer_bind     complement(1085..1104)
                     /label=SMP2433
     primer_bind     complement(1100..1118)
                     /label=SMP3388
     primer_bind     1143..1163
                     /label=SMP2316
     primer_bind     1217..1236
                     /label=SMP1001
     primer_bind     1263..1282
                     /label=SMP1325
     primer_bind     complement(1273..1293)
                     /label=SMP2969
     primer_bind     complement(1330..1350)
                     /label=SMP2971
     primer_bind     complement(1333..1349)
                     /label=SMP2578
     primer_bind     complement(1363..1376)
                     /label=SMP2969
     primer_bind     1376..1394
                     /label=SMP444
     primer_bind     1386..1410
                     /label=SMP1666
     primer_bind     1440..1457
                     /label=SMP3040
     primer_bind     1440..1457
                     /label=SMP3104
     primer_bind     1445..1464
                     /label=SMP1330
     primer_bind     1472..1491
                     /label=SMP437
     primer_bind     complement(1472..1491)
                     /label=SMP471
     primer_bind     1523..1542
                     /label=SMP364
     primer_bind     1572..1604
                     /label=SMP295
     primer_bind     complement(1572..1604)
                     /label=SMP296
     primer_bind     1595..1613
                     /label=EGFP-C-F-31
     primer_bind     1604..1620
                     /label=EGFP-for
     primer_bind     1610..1628
                     /label=SMP564
     primer_bind     1622..1643
                     /label=SMP3300
     primer_bind     1626..1643
                     /label=EGFP-C-for
     primer_bind     complement(1670..1707)
                     /label=Vector.REV (1)
     primer_bind     1683..1722
                     /label=Fragment.FORa
     CDS             1698..1817
                     /codon_start=1
                     /label=ATG13-IDR (191-230)
                     /note="ATG13"
                     /translation="INLAFMSTRQFERTPPIMGIIIDHFVDRPYPSSSPMHPCN"
     misc_feature    1779..1796
                     /label=SMP2547
     primer_bind     1779..1796
                     /label=SMP2547
     misc_feature    1790..1825
                     /label=rev
     misc_feature    1811..1841
                     /label=fw
     primer_bind     complement(1818..1837)
                     /label=Fragment.REV
     primer_bind     1823..1842
                     /label=Vector.FOR
     primer_bind     complement(1862..1881)
                     /label=SMP_3644
     primer_bind     2147..2167
                     /label=SMP_646
     primer_bind     complement(2147..2167)
                     /label=SMP_654
     promoter        2148..2252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2253..3113
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3284..3872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3664..3681
                     /label=SMP_4077
     promoter        4116..4193
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4194..5276
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4461..4478)
                     /label=SMP_4079
     primer_bind     5257..5274
                     /label=SMP_2212
     primer_bind     complement(5259..5276)
                     /label=SMP_4078
     protein_bind    5289..5310
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5325..5355
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5363..5379
                     /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     complement(5371..5396)
                     /label=Vector.REV
     primer_bind     5382..5401
                     /label=SMP_993
     primer_bind     5387..5403
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5387..5403
                     /label=SMP_1770
     CDS             join(5399..5845,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGITLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5419..5435)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5420..5435)
                     /label=SMP_1771
     primer_bind     complement(5423..5444)
                     /label=SMP_469
     primer_bind     complement(5459..5479)
                     /label=SMP_3886
     primer_bind     complement(5465..5485)
                     /label=SMP_3888
     primer_bind     complement(5473..5492)
                     /label=SMP_3890
ORIGIN
        1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg gcatccccta tactaggtta ttggaaaatt aagggccttg
      301 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
      361 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
      421 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
      481 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
      541 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
      601 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
      661 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
      721 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
      781 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
      841 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
      901 accatcctcc aaaatcggat gattacgata tcccaacgac cgaaaacctg tattttcagg
      961 gtggtaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1021 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1081 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1141 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     1201 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1261 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1321 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1381 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     1441 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     1501 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1561 acaaccacta cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1621 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1681 acaagggtac cggtggtatc aatctggcct tcatgtcaac cagacagttt gagcgtactc
     1741 cccctattat gggaatcatt atcgaccact tcgtggatag accatacccc agtagctccc
     1801 ctatgcatcc atgcaactga cccgggtgac tcgagcggcc gcatcgtgac tgactgacga
     1861 tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     1921 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     1981 cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt
     2041 ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2221 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2281 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2341 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2401 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2461 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     2521 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2581 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2641 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2701 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2761 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     2821 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2881 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2941 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3001 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3061 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3421 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3901 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3961 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4021 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4081 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct
     4141 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     4201 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     4261 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     4321 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     4381 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     4441 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     4501 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     4561 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     4621 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     4681 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     4741 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     4801 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     4861 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     4921 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     4981 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     5041 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     5101 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     5161 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     5221 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     5281 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     5341 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     5401 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     5461 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     5521 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt
     5581 tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga
     5641 ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta
     5701 caccaacgta acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac
     5761 gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg
     5821 aattattttt gatggcgttg gaatt
//
