LOCUS       Exported                6631 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 6631)
  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..6631
                     /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=mEGFP
                     /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(1670..1707)
                     /label=Vector.REV (1)
     primer_bind     1683..1722
                     /label=Fragment.FORa
     CDS             1698..2603
                     /codon_start=1
                     /label=ATG13-IDR delta206-230 (191-205, 231-517)
                     /note="ATG13"
                     /translation="INLAFMSTRQFERTPYRTAGEDTGVIYPSVEDSQEVCTTSFSTSP
                     PSQLSSSRLSYQPAALGVGSADLAYPVVFAAGLNATHPHQLMVPGKEGGVPLAPNQPVH
                     GTQADQERLATCTPSDRTHCAATPSSSEDTETVSNSSEGRASPHDVLETIFVRKVGAFV
                     NKPINQVTLTSLDIPFAMFAPKNLELEDTDPMVNPPDSPETESPLQGSLHSDGSSGGSS
                     GNTHDDFVMIDFKPAFSKDDILPMDLGTFYREFQNPPQLSSLSIDIGAQSMAEDLDSLP
                     EKLAVHEKNVREFDAFVETLQ"
     primer_bind     complement(1841..1857)
                     /label=SMP2554
     primer_bind     complement(2597..2623)
                     /label=Fragment.REV
     primer_bind     2609..2628
                     /label=Vector.FOR
     primer_bind     complement(2648..2667)
                     /label=SMP_3644
     primer_bind     2933..2953
                     /label=SMP_646
     primer_bind     complement(2933..2953)
                     /label=SMP_654
     promoter        2934..3038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3039..3899
                     /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      4070..4658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4450..4467
                     /label=SMP_4077
     promoter        4902..4979
                     /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             4980..6062
                     /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(5247..5264)
                     /label=SMP_4079
     primer_bind     6043..6060
                     /label=SMP_2212
     primer_bind     complement(6045..6062)
                     /label=SMP_4078
     protein_bind    6075..6096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6111..6141
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6149..6165
                     /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(6157..6182)
                     /label=Vector.REV
     primer_bind     6168..6187
                     /label=SMP_993
     primer_bind     6173..6189
                     /label=SMP_1770
     CDS             join(6185..6631,1..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(6206..6221)
                     /label=SMP_1771
     primer_bind     complement(6209..6230)
                     /label=SMP_469
     primer_bind     complement(6245..6265)
                     /label=SMP_3886
     primer_bind     complement(6251..6271)
                     /label=SMP_3888
     primer_bind     complement(6259..6278)
                     /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 cctatcgtac agccggtgaa gacaccggcg tgatctaccc cagtgtcgag gatagccagg
     1801 aagtgtgtac cacttccttc tctacctcac caccctcaca gctctcttca agtcgcctga
     1861 gttatcagcc tgccgctctc ggagtcggta gcgcagacct ggcctaccca gtggtcttcg
     1921 cagcaggcct caacgcaacc cacccacatc agctgatggt gccaggaaag gagggtggag
     1981 tcccactggc tcctaatcag ccagtgcacg gtactcaggc tgaccaggaa aggctggcaa
     2041 cttgcacacc atccgataga actcattgtg ctgcaacacc cagctcctct gaggacaccg
     2101 aaactgtgag caactcaagt gagggtcgcg cctccccaca cgatgtcctg gaaaccatct
     2161 tcgtgagaaa agtcggcgct tttgtgaaca agcccattaa tcaggtcaca ctcacctctc
     2221 tggacatccc tttcgctatg tttgcaccaa agaacctcga gctggaagac actgatccca
     2281 tggtgaatcc tccagactca cccgagacag aaagtcctct ccagggcagt ctgcacagcg
     2341 atggaagctc cggtggctct tcaggcaaca cacatgacga tttcgtcatg attgacttca
     2401 aacccgcttt ttccaaggac gatatcctcc ctatggatct gggaaccttc tacagggagt
     2461 ttcagaatcc ccctcagctc agtagcctga gcattgacat cggtgcccag tccatggctg
     2521 aggacctcga ttctctgcct gaaaaactgg ctgtgcacga gaagaacgtc agagaattcg
     2581 acgcatttgt ggagaccctg cagtgacccg ggtgactcga gcggccgcat cgtgactgac
     2641 tgacgatctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2701 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2761 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     2821 gagtgtataa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     2881 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     2941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3001 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3061 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3121 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3181 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3241 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     3301 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3361 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3421 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3481 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3541 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     3601 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3661 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3721 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3781 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3841 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3901 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3961 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4201 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4741 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4801 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     4861 tccttacgca tctgtgcggt atttcacacc gcataaattc cgacaccatc gaatggtgca
     4921 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     4981 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     5041 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     5101 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     5161 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     5221 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     5281 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     5341 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     5401 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     5461 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     5521 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     5581 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
     5641 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
     5701 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
     5761 ggctgcgcgt tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat
     5821 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
     5881 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
     5941 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
     6001 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6061 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6121 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6181 agctatgacc atgattacgg attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     6241 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     6301 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     6361 gcgctttgcc tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct
     6421 tcctgaggcc gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc
     6481 catctacacc aacgtaacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa
     6541 tccgacgggt tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca
     6601 gacgcgaatt atttttgatg gcgttggaat t
//
