pGEX4T-1-AUREO1c

LOCUS       pGEX4T-1-AUREO1c          5947 bp DNA     circular SYN 04-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGEX 4T-1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5947)
  AUTHORS   Huanhuan
  TITLE     Direct Submission
  JOURNAL   Exported Sep 26, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    183..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"
     misc_feature    916..957
                     /label=HZ-1387 F
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             936..1913
                     /codon_start=1
                     /label=AUREO1c Huan template
                     /translation="ADQAKANPSGPSNTAPLFQMSDPANPSSTLDLDDLFNDIYYMDLP
                     NGSNMDSDPTPVDVPSSNGNGHDGASKKRSADDFDGDSLDGTGNKDLTEQQKLERRERN
                     REHAKRSRLRKKFLLESLQEQIHGLEEQLDGLKSAIKKELPQQAEQIITRICGDKEKFT
                     PLPMPSGFGPVKTLMEPDFRLMSALSGSQQNFAISDPTLPDNPIVYVSQGFLDLTGYTL
                     DQVLGRNCRFLQGPGTDQSAVEVIRKGITEGVDTSVCLLNYKADGTPFWNQFFVASLRD
                     AENNVVNHVGVQCEVSKAVVEKHMGEQKKAAAEAAKARPVTTTSS"
     misc_feature    1184..1203
                     /label=HZ-197 靶标序列
     primer_bind     complement(1895..1936)
                     /label=HZ-1388 R
     promoter        2250..2354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2355..3215
                     /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      3386..3974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4218..4295
                     /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             4296..5378
                     /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"
     protein_bind    5391..5412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5427..5457
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5465..5481
                     /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     5489..5505
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(5501..5947,1..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(5521..5537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 gttccgcgtg gatccgcgga ccaagccaag gccaatccat
      961 ccggaccgtc gaatacggca cctttatttc aaatgtctga tccggcgaat cccagttcca
     1021 ctctggatct cgacgacttg ttcaacgata tctattacat ggatttaccg aacggcagca
     1081 atatggattc ggatcctacg cccgtggacg tgcccagcag caacgggaac ggccacgacg
     1141 gtgcgagtaa gaaacgcagt gcggatgatt tcgacgggga cagtttggac gggaccggca
     1201 ataaggattt gacggagcag cagaaattgg agcgccgtga gcgcaaccgc gaacacgcca
     1261 aacgtagccg tctccgcaaa aagtttctac tcgaatcgct gcaggaacag attcacgggt
     1321 tggaggaaca gttggatggt ttgaaatcgg caattaaaaa ggaattacca caacaagcgg
     1381 aacaaattat taccaggatt tgtggggata aggaaaagtt tacgccgctg cccatgccga
     1441 gtggatttgg acccgtgaag acgctcatgg agccggactt tcggctcatg tcggctcttt
     1501 ccggttcgca gcaaaatttc gctatttcgg atccaactct tcccgataat cctattgttt
     1561 atgttagtca aggatttctg gatttgaccg gatacacgct ggatcaggtc ttgggtcgca
     1621 attgtcgatt tctgcaggga ccaggaaccg accagagtgc agtggaagtc attcgcaagg
     1681 gaattactga aggagtggac acgtccgtgt gtttgctgaa ttacaaagcc gacggcacac
     1741 ccttttggaa ccaattcttt gtggcctctc tacgagacgc cgaaaacaac gtggtcaatc
     1801 acgtcggggt tcagtgtgaa gtgagcaagg ccgtggtcga aaaacacatg ggggaacaaa
     1861 agaaggccgc agcagaagcc gccaaagccc gacccgtgac cactactagt agttgagaat
     1921 tcccgggtcg actcgagcgg ccgcatcgtg actgactgac gatctgcctc gcgcgtttcg
     1981 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2041 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2101 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa
     2161 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2221 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2281 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2341 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2401 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2461 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2521 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2581 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2641 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2701 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2761 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2821 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2881 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2941 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3001 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3061 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3121 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3301 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3361 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3421 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3481 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3541 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3601 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3661 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3721 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3781 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3841 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3901 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3961 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4021 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4081 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4141 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4201 cacaccgcat aaattccgac accatcgaat ggtgcaaaac ctttcgcggt atggcatgat
     4261 agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg
     4321 tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc
     4381 acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc
     4441 ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct
     4501 ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc
     4561 aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag
     4621 cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg
     4681 atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg
     4741 atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac
     4801 tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat
     4861 taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc actcgcaatc
     4921 aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt tttcaacaaa
     4981 ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc aacgatcaga
     5041 tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt gcggatatct
     5101 cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg ttaaccacca
     5161 tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg caactctctc
     5221 agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa agaaaaacca
     5281 ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     5341 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     5401 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     5461 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacggattc
     5521 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5581 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5641 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc tttgcctggt ttccggcacc
     5701 agaagcggtg ccggaaagct ggctggagtg cgatcttcct gaggccgata ctgtcgtcgt
     5761 cccctcaaac tggcagatgc acggttacga tgcgcccatc tacaccaacg taacctatcc
     5821 cattacggtc aatccgccgt ttgttcccac ggagaatccg acgggttgtt actcgctcac
     5881 atttaatgtt gatgaaagct ggctacagga aggccagacg cgaattattt ttgatggcgt
     5941 tggaatt
//