LOCUS       sequence-13828-e        7040 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7040)
  TITLE     Koloteva-Levine et al
REFERENCE   2  (bases 1 to 7040)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pTH385-GST-TEV-SUP35
FEATURES             Location/Qualifiers
     source          1..7040
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..941
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             951..971
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3090..3112)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3257..3275)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3343..3447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3448..3516,3517..4308)
                     /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"
     primer_bind     complement(3666..3685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4479..5067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4968..4987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5221..5238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5311..5388
                     /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             5389..6471
                     /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
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5408..5427)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6484..6505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6520..6525,6526..6543,6544..6550)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6558..6574
                     /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     6582..6598
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6582..6598
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6594..7040,1..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(6614..6631)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6614..6630)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6623..6645)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
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 tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc gaaaatttgt
      961 attttcaagg tatgtcggat tcaaaccaag gcaacaatca gcaaaactac cagcaataca
     1021 gccagaacgg taaccaacaa caaggtaaca acagatacca aggttatcaa gcttacaatg
     1081 ctcaagccca acctgcaggt gggtactacc aaaattacca aggttattct gggtaccaac
     1141 aaggtggcta tcaacagtac aatcccgacg ccggttacca gcaacagtat aatcctcaag
     1201 gaggctatca acagtacaat cctcaaggcg gttatcagca gcaattcaat ccacaaggtg
     1261 gccgtggaaa ttacaaaaac ttcaactaca ataacaattt gcaaggatat caagctggtt
     1321 tccaaccaca gtctcaaggt atgtctttga acgactttca aaagcaacaa aagcaggccg
     1381 ctcccaaacc aaagaagact ttgaagcttg tctccagttc cggtatcaag ttggccaatg
     1441 ctaccaagaa ggttggcaca aaacctgccg aatctgataa gaaagaggaa gagaagtctg
     1501 ctgaaaccaa agaaccaact aaagagccaa caaaggtcga agaaccagtt aaaaaggagg
     1561 agaaaccagt ccagactgaa gaaaagacgg aggaaaaatc ggaacttcca aaggtagaag
     1621 accttaaaat ctctgaatca acacataata ccaacaatgc caatgttacc agtgctgatg
     1681 ccttgatcaa ggaacaggaa gaagaagtgg atgacgaagt tgttaacgat atgtttggtg
     1741 gtaaagatca cgtttcttta attttcatgg gtcatgttga tgccggtaaa tctactatgg
     1801 gtggtaatct actatacttg actggctctg tggataagag aactattgag aaatatgaaa
     1861 gagaagccaa ggatgcaggc agacaaggtt ggtacttgtc atgggtcatg gataccaaca
     1921 aagaagaaag aaatgatggt aagactatcg aagttggtaa ggcctacttt gaaactgaaa
     1981 aaaggcgtta taccatattg gatgctcctg gtcataaaat gtacgtttcc gagatgatcg
     2041 gtggtgcttc tcaagctgat gttggtgttt tggtcatttc cgccagaaag ggtgagtacg
     2101 aaaccggttt tgagagaggt ggtcaaactc gtgaacacgc cctattggcc aagacccaag
     2161 gtgttaataa gatggttgtc gtcgtaaata agatggatga cccaaccgtt aactggtcta
     2221 aggaacgtta cgaccaatgt gtgagtaatg tcagcaattt cttgagagca attggttaca
     2281 acattaagac agacgttgta tttatgccag tatccggcta cagtggtgca aatttgaaag
     2341 atcacgtaga tccaaaagaa tgcccatggt acaccggccc aactctgtta gaatatctgg
     2401 atacaatgaa ccacgtcgac cgtcacatca atgctccatt catgttgcct attgccgcta
     2461 agatgaagga tctaggtacc atcgttgaag gtaaaattga atccggtcat atcaaaaagg
     2521 gtcaatccac cctactgatg cctaacaaaa ccgctgtgga aattcaaaat atttacaacg
     2581 aaactgaaaa tgaagttgat atggctatgt gtggtgagca agttaaacta agaatcaaag
     2641 gtgttgaaga agaagacatt tcaccaggtt ttgtactaac atcgccaaag aaccctatca
     2701 agagtgttac caagtttgta gctcaaattg ctattgtaga attaaaatct atcatagcag
     2761 ccggtttttc atgtgttatg catgttcata cagcaattga agaggtacat attgttaagt
     2821 tattgcacaa attagaaaag ggtaccaacc gtaagtcaaa gaaaccacct gcttttgcta
     2881 agaagggtat gaaggtcatc gctgttttag aaactgaagc tccagtttgt gtggaaactt
     2941 accaagatta ccctcaatta ggtagattca ctttgagaga tcaaggtacc acaatagcaa
     3001 ttggtaaaat tgttaaaatt gccgagtaag cggccgcatc gtgactgact gacgatctgc
     3061 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     3121 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     3181 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtataat
     3241 tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3301 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3361 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3421 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3481 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3541 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3601 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3661 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     3721 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3781 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3841 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3901 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3961 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4021 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4081 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4141 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4201 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4261 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4321 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4381 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4441 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4501 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4561 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4621 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4681 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4741 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4801 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4861 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4921 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4981 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5041 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5101 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5161 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5221 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5281 ctgtgcggta tttcacaccg cataaattcc gacaccatcg aatggtgcaa aacctttcgc
     5341 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     5401 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     5461 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     5521 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     5581 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     5641 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     5701 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     5761 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     5821 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     5881 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     5941 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     6001 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     6061 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     6121 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     6181 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     6241 ccgtcaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     6301 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     6361 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6421 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6481 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6541 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     6601 tgattacgga ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     6661 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     6721 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgctttgcct
     6781 ggtttccggc accagaagcg gtgccggaaa gctggctgga gtgcgatctt cctgaggccg
     6841 atactgtcgt cgtcccctca aactggcaga tgcacggtta cgatgcgccc atctacacca
     6901 acgtaaccta tcccattacg gtcaatccgc cgtttgttcc cacggagaat ccgacgggtt
     6961 gttactcgct cacatttaat gttgatgaaa gctggctaca ggaaggccag acgcgaatta
     7021 tttttgatgg cgttggaatt
//
