LOCUS       sequence-247073-        6748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6748)
  AUTHORS   Widlund PO, Podolski M, Reber S, Alper J, Storch M, Hyman AA, Howard
            J, Drechsel DN
  TITLE     One-step purification of assembly competent tubulin from diverse 
            eukaryotic sources.
  JOURNAL   Mol Biol Cell. 2012 Sep 19.
  PUBMED    22993214
REFERENCE   2  (bases 1 to 6748)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2012 Sep 19."
COMMENT     Sequence Label: pGEX-6P-1 Stu2 1-590
FEATURES             Location/Qualifiers
     source          1..6748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(102..107,108..123,124..130)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    138..154
                     /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             177..830
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     788..810
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             837..860
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(2717..2739)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2884..2902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2970..3074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3075..3143,3144..3935)
                     /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(3293..3312)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4106..4694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4595..4614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4848..4865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4938..5015
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5016..6098
                     /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(5035..5054)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6147..6152,6153..6170,6171..6177)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6185..6201
                     /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     6209..6225
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6209..6225
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6221..6748
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6241..6258)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6241..6257)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6250..6272)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
       61 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatcggc
      121 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca gtattcatgt
      181 cccctatact aggttattgg aaaattaagg gccttgtgca acccactcga cttcttttgg
      241 aatatcttga agaaaaatat gaagagcatt tgtatgagcg cgatgaaggt gataaatggc
      301 gaaacaaaaa gtttgaattg ggtttggagt ttcccaatct tccttattat attgatggtg
      361 atgttaaatt aacacagtct atggccatca tacgttatat agctgacaag cacaacatgt
      421 tgggtggttg tccaaaagag cgtgcagaga tttcaatgct tgaaggagcg gttttggata
      481 ttagatacgg tgtttcgaga attgcatata gtaaagactt tgaaactctc aaagttgatt
      541 ttcttagcaa gctacctgaa atgctgaaaa tgttcgaaga tcgtttatgt cataaaacat
      601 atttaaatgg tgatcatgta acccatcctg acttcatgtt gtatgacgct cttgatgttg
      661 ttttatacat ggacccaatg tgcctggatg cgttcccaaa attagtttgt tttaaaaaac
      721 gtattgaagc tatcccacaa attgataagt acttgaaatc cagcaagtat atagcatggc
      781 ctttgcaggg ctggcaagcc acgtttggtg gtggcgacca tcctccaaaa tcggatctgg
      841 aagttctgtt ccaggggccc ctgggatccc cggaattcat gtcaggagaa gaagaagtag
      901 attatacaac gttaccccta gaagagcgat taacgtacaa gttatggaag gcacgtttag
      961 aagcttataa ggagttaaac cagcttttta gaaactccgt tggcgacatc agtagggatg
     1021 acaacataca aatttactgg agggatccta cattatttgc acaatatata actgattcta
     1081 atgtggtcgc tcaagaacaa gccattgttg ccttgaatag tttaatagat gcctttgcgt
     1141 catcgtcttt gaaaaatgca cataatatta ctttaatatc aacatggaca ccattgcttg
     1201 tggaaaaggg cttaacgtcc tcaagggcta caacgaagac gcagtcgatg agctgtatac
     1261 tctcactatg tggtcttgat acatctatta cgcaatcggt ggaattggta ataccctttt
     1321 tcgagaaaaa actgccaaaa ttaattgctg cggcagccaa ttgtgtttat gagttgatgg
     1381 ctgcattcgg gctcactaat gtgaacgttc aaacgtttct accagaatta ttaaagcacg
     1441 tacctcaact agccggtcat ggtgatagaa acgttcgatc acaaacgatg aatttgattg
     1501 tcgagatcta taaggttacg ggcaacaata gcgacttgtt agaggagata ctgtttaaaa
     1561 aattaaaacc aattcaagtg aaagatcttc ataaattgtt tgcaaaagtt ggtgacgagc
     1621 catcgtcatc gaaaatgtta ttcgagtggg agaaaagaga attggaaaag aaaagaagcc
     1681 aagaggagga agccaggaaa agaaagtcca tattatcgaa tgatgagggg gaatatcaga
     1741 tagataagga tggagataca ttgatgggga tggaaacgga tatgccacca tcaaaacagc
     1801 aaagtggagt tcaaatagac accttcagta tgctaccgga ggagacgatc ctcgataaac
     1861 tgcctaagga cttccaggaa aggataacgt cgtccaaatg gaaagataga gtggaagcgc
     1921 tggaagaatt ttgggatagt gttctatcgc agactaagaa gttaaaaagc acctcgcaaa
     1981 attattccaa cttattgggg atatatggcc atattatcca gaaagatgca aacattcaag
     2041 ctgttgcact agccgctcaa tcagtagaat taatctgcga caaactaaaa acaccaggct
     2101 tttccaagga ttatgtttca ctagtcttca ctcccttgtt agacaggaca aaggaaaaga
     2161 aaccttctgt gatagaggcc attaggaagg cattattaac catctgcaag tattacgatc
     2221 cattagcctc cagtggacgc aatgaagata tgttgaaaga tattttggag catatgaagc
     2281 acaagacacc tcaaatcaga atggagtgta cgcaattgtt taatgcaagc atgaaagaag
     2341 aaaaagatgg atactcgacc ctgcaaagat acttgaagga tgaagtggta ccaattgtga
     2401 ttcagatcgt caacgacacc caacctgcta taagaactat tggatttgag agtttcgcca
     2461 ttctaattaa aatatttggc atgaatacct ttgtgaaaac attagagcac ttggataatt
     2521 tgaaaaggaa aaaaattgaa gaaacggtca aaactctacc taatttcagt attgcaagtg
     2581 gtagtaccca tagcactatt gaaacgaata aacaaactgg gccaatggag aataaatttc
     2641 tcctgaaata actcgagcgg ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg
     2701 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2761 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2821 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa
     2881 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     3061 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     3121 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     3181 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     3241 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     3301 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     3361 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     3421 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3481 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3541 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3601 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     3661 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3721 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3781 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3841 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3901 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3961 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4021 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     4261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4861 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4921 cacaccgcat aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat
     4981 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     5041 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     5101 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     5161 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     5221 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     5281 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     5341 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     5401 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     5461 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     5521 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     5581 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     5641 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     5701 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     5761 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     5821 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     5881 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     5941 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     6001 cgctagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6061 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt
     6121 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     6181 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc
     6241 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     6301 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     6361 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
     6421 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
     6481 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg tgacctatcc
     6541 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
     6601 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
     6661 tggaattagc ttatcgactg cacggtgcac caatgcttct ggcgtcaggc agccatcgga
     6721 agctgtggta tggctgtgca ggtcgtaa
//
