pGEX-2T-NXT3

LOCUS       pGEX-2T-NXT3               6255 bp DNA     circular SYN 10-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6255)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 16, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        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"
     CDS             918..938
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             942..2246
                     /codon_start=1
                     /label=NXT3 Codon
                     /translation="MTAENATLLEPVLGKDEIGWMFVQEYYTYLNKEPNRLHCFYTKKS
                     TLIHGDEGESISLCHGQQEIHNKILDLDFQNCKVLISNVDSLASSNGGIVIQVLGEMSN
                     KGKLSRKFAQTFFLAEQPNGYFVLNDIFRFLREDVEEEEESPDAVEKEKKDVASEPYVN
                     GVQSQEHLPSAKEEGHYQDPAATENNFATAALISNETDSLNQATLAVPEEPVIQVTEAS
                     VPSFVSQQENQLQDEALTSNSKNADAIGASDANVATAPKSWADLIARNHPDVKSQASVS
                     STASTTGQTVKGVNADQTQQPTAPYTQSNELLETSVFVKNIPPETSDVSLKSAMSIFGP
                     VKAIEFARRKGTAYVDFVNHECVQLALNKKTLQINNATLNIEERRRLFSGKFNKSGDKK
                     SNDNYNGMKRNFRKGNRGAFDGRSKEVTTSKKQNN"
     promoter        2558..2662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2663..3523
                     /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      3694..4282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        4526..4603
                     /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             4604..5686
                     /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    5699..5720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5735..5765
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5773..5789
                     /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     5797..5813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(5809..6255,1..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(5829..5845)
                     /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 atcggatgaa aacctgtatt ttcagggatc catgaccgcc gaaaatgcaa
      961 ccctgctgga accggtgctg ggcaaagatg aaattggctg gatgtttgtt caggaatatt
     1021 atacctatct gaacaaagaa ccgaatcgcc tgcattgctt ttataccaaa aaatcaaccc
     1081 tgattcacgg tgacgaaggc gaaagcatta gtctgtgcca cggtcagcag gaaattcata
     1141 ataagattct ggatctggat ttccagaatt gtaaagttct gattagtaac gtggatagtc
     1201 tggcaagcag caatggtggt attgttattc aggtgctggg tgaaatgagc aataagggta
     1261 aactgagccg caaatttgca cagacctttt tcctggcaga acagccgaat ggctattttg
     1321 tgctgaatga tattttccgc tttctgcgcg aagatgtgga agaagaagaa gagagcccgg
     1381 atgcagttga aaaagaaaag aaagatgtgg caagtgaacc gtatgtgaat ggtgtgcaga
     1441 gtcaggaaca tctgccgagt gccaaagaag aaggtcatta tcaggaccct gccgccaccg
     1501 aaaataattt tgccaccgca gcactgatta gcaatgaaac cgatagtctg aatcaggcca
     1561 ccctggccgt gccggaagaa ccggttattc aggttaccga agccagcgtt ccgagttttg
     1621 tgagtcagca ggaaaatcag ctgcaggatg aagcactgac cagcaatagt aaaaatgccg
     1681 atgccattgg tgccagtgat gcaaatgttg ccaccgcccc gaaaagttgg gcagatctga
     1741 ttgcccgcaa tcatccggat gttaaaagtc aggccagcgt gagcagcacc gcaagtacca
     1801 ccggtcagac cgttaaaggt gttaatgcag atcagaccca gcagccgacc gcaccgtata
     1861 cccagagtaa tgaactgctg gaaaccagtg tgtttgtgaa aaatattccg ccggaaacca
     1921 gcgatgtgag tctgaaaagc gcaatgagca tttttggccc ggtgaaagcc attgaatttg
     1981 cacgccgtaa aggtaccgcc tatgtggatt ttgtgaatca tgaatgcgtg cagctggccc
     2041 tgaataagaa aaccctgcag attaataatg caaccttaaa tatcgaggag cgtcgccgcc
     2101 tgtttagcgg caaattcaat aagagtggtg acaaaaaatc caatgataat tacaacggca
     2161 tgaaacgcaa ttttcgtaaa ggcaatcgtg gtgcctttga tggccgtagc aaagaagtga
     2221 ccaccagcaa aaaacagaat aattgagaat tcatcgtgac tgactgacga tctgcctcgc
     2281 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2341 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2401 cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat agcggagtgt ataattcttg
     2461 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     2521 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2881 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     2941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3181 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     3241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3841 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4441 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4501 cggtatttca caccgcataa attccgacac catcgaatgg tgcaaaacct ttcgcggtat
     4561 ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt
     4621 atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca
     4681 ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa
     4741 ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt
     4801 tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg
     4861 cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc
     4921 ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga tcattaacta
     4981 tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt
     5041 atttcttgat gtctctgacc agacacccat caacagtatt attttctccc atgaagacgg
     5101 tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg cgctgttagc
     5161 gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata aatatctcac
     5221 tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca tgtccggttt
     5281 tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc tggttgccaa
     5341 cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc gcgttggtgc
     5401 ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata tcccgccgtt
     5461 aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca
     5521 actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag
     5581 aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5821 acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5881 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5941 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt
     6001 ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg atcttcctga ggccgatact
     6061 gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg cgcccatcta caccaacgta
     6121 acctatccca ttacggtcaa tccgccgttt gttcccacgg agaatccgac gggttgttac
     6181 tcgctcacat ttaatgttga tgaaagctgg ctacaggaag gccagacgcg aattattttt
     6241 gatggcgttg gaatt
//