LOCUS       Exported                6664 bp ds-DNA     circular SYN 04-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6664)
  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..6664
                     /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
     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
     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(1668..1712)
                     /label=SMP5470
     primer_bind     1693..1742
                     /label=SMP5469
     CDS             1698..2636
                     /codon_start=1
                     /label=ATG13-IDR_delta191-204 (205-517aa)
                     /note="ATG13"
                     /translation="PPIMGIIIDHFVDRPYPSSSPMHPCNYRTAGEDTGVIYPSVEDSQ
                     EVCTTSFSTSPPSQLSSSRLSYQPAALGVGSADLAYPVVFAAGLNATHPHQLMVPGKEG
                     GVPLAPNQPVHGTQADQERLATCTPSDRTHCAATPSSSEDTETVSNSSEGRASPHDVLE
                     TIFVRKVGAFVNKPINQVTLTSLDIPFAMFAPKNLELEDTDPMVNPPDSPETESPLQGS
                     LHSDGSSGGSSGNTHDDFVMIDFKPAFSKDDILPMDLGTFYREFQNPPQLSSLSIDIGA
                     QSMAEDLDSLPEKLAVHEKNVREFDAFVETLQ"
     primer_bind     complement(1874..1890)
                     /label=SMP2554
     primer_bind     complement(2630..2656)
                     /label=Fragment.REV
     primer_bind     2642..2661
                     /label=Vector.FOR
     primer_bind     complement(2681..2700)
                     /label=SMP_3644
     primer_bind     2966..2986
                     /label=SMP_646
     primer_bind     complement(2966..2986)
                     /label=SMP_654
     promoter        2967..3071
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3072..3932
                     /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      4103..4691
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4483..4500
                     /label=SMP_4077
     promoter        4935..5012
                     /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             5013..6095
                     /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(5280..5297)
                     /label=SMP_4079
     primer_bind     6076..6093
                     /label=SMP_2212
     primer_bind     complement(6078..6095)
                     /label=SMP_4078
     protein_bind    6108..6129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6144..6174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6182..6198
                     /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(6190..6215)
                     /label=Vector.REV
     primer_bind     6201..6220
                     /label=SMP_993
     primer_bind     6206..6222
                     /label=SMP_1770
     CDS             join(6218..6664,1..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(6239..6254)
                     /label=SMP_1771
     primer_bind     complement(6242..6263)
                     /label=SMP_469
     primer_bind     complement(6278..6298)
                     /label=SMP_3886
     primer_bind     complement(6284..6304)
                     /label=SMP_3888
     primer_bind     complement(6292..6311)
                     /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 cggtggtccc cctattatgg gaatcattat cgaccacttc gtggatagac
     1741 cataccccag tagctcccct atgcatccat gcaactatcg tacagccggt gaagacaccg
     1801 gcgtgatcta ccccagtgtc gaggatagcc aggaagtgtg taccacttcc ttctctacct
     1861 caccaccctc acagctctct tcaagtcgcc tgagttatca gcctgccgct ctcggagtcg
     1921 gtagcgcaga cctggcctac ccagtggtct tcgcagcagg cctcaacgca acccacccac
     1981 atcagctgat ggtgccagga aaggagggtg gagtcccact ggctcctaat cagccagtgc
     2041 acggtactca ggctgaccag gaaaggctgg caacttgcac accatccgat agaactcatt
     2101 gtgctgcaac acccagctcc tctgaggaca ccgaaactgt gagcaactca agtgagggtc
     2161 gcgcctcccc acacgatgtc ctggaaacca tcttcgtgag aaaagtcggc gcttttgtga
     2221 acaagcccat taatcaggtc acactcacct ctctggacat ccctttcgct atgtttgcac
     2281 caaagaacct cgagctggaa gacactgatc ccatggtgaa tcctccagac tcacccgaga
     2341 cagaaagtcc tctccagggc agtctgcaca gcgatggaag ctccggtggc tcttcaggca
     2401 acacacatga cgatttcgtc atgattgact tcaaacccgc tttttccaag gacgatatcc
     2461 tccctatgga tctgggaacc ttctacaggg agtttcagaa tccccctcag ctcagtagcc
     2521 tgagcattga catcggtgcc cagtccatgg ctgaggacct cgattctctg cctgaaaaac
     2581 tggctgtgca cgagaagaac gtcagagaat tcgacgcatt tgtggagacc ctgcagtgac
     2641 ccgggtgact cgagcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg
     2701 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2761 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2821 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg
     2881 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2941 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3001 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3061 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3121 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3181 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3241 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3301 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3361 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3421 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3481 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3541 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3601 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3661 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3721 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3781 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3841 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3901 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3961 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4021 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4081 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4141 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4201 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4261 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4321 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4381 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4441 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4501 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4561 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4621 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4681 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4741 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4801 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4861 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4921 accgcataaa ttccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
     4981 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     5041 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     5101 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     5161 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     5221 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     5281 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5341 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5401 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5461 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5521 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5581 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5641 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5701 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     5761 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     5821 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     5881 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     5941 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     6001 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     6061 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     6121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
     6241 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6301 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6361 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga
     6421 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
     6481 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtaa cctatcccat
     6541 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
     6601 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg
     6661 aatt
//
