LOCUS       sequence-358705-        5702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli optimized expression of GST-fusion of human NDFIP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5702)
  TITLE     Haas lab plasmids 2021
REFERENCE   2  (bases 1 to 5702)
  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"
COMMENT     Sequence Label: pGEX4T1-NDFIP2
FEATURES             Location/Qualifiers
     source          1..5702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(555..560,561..578,579..585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    593..609
                     /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     617..633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     617..633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             629..1156
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(649..666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(649..665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(658..680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1258..1263,1264..1279,1280..1286)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1294..1310
                     /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             1333..1986
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1944..1966
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1993..2010
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2827..2849)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2994..3012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3080..3184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3185..3253,3254..4045)
                     /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(3403..3422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4216..4804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4705..4724
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4958..4975
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5048..5125
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5126..5702,1..506)
                     /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(5145..5164)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
       61 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      121 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      181 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
      241 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
      301 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
      361 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
      421 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      481 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
      541 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      601 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
      661 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
      721 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
      781 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
      841 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
      901 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
      961 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     1021 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt
     1081 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     1141 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1201 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     1261 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     1321 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
     1381 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
     1441 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
     1501 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
     1561 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
     1621 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
     1681 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
     1741 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
     1801 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
     1861 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
     1921 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
     1981 ccaaaatcgg atctggttcc gcgtggatcc ccggaattcc cgggtcgaca aatggatcac
     2041 caccagccgg ggactgggcg ctaccaggtg cttcttaatg aagaggataa ctcagaatca
     2101 tcggctatag agcagccacc tacttcaaac ccagcaccgc agattgtgca ggctgtgtct
     2161 tcagcaccag cacttgaaac tgactcttcc cctccaccat atagtagtat tactgtggaa
     2221 gtacctacaa cttcagatac agaagtttac ggtgagtttt atcccgtgcc acctccctat
     2281 agcgttgcta cctctcttcc tacatacgat gaagctgaga aggctaaagc tgctgcaatg
     2341 gcagctgcag cagcagaaac atctcaaaga attcaggagg aagagtgtcc accaagagat
     2401 gacttcagtg atgcagacca gctcagagtg gggaatgatg gcattttcat gctggcattt
     2461 ttcatggcat ttattttcaa ctggcttgga ttttgtttat ccttctgtat caccaatacc
     2521 atagctggaa ggtatggtgc tatctgcgga tttggccttt ccttgatcaa atggatcctt
     2581 attgtcaggt tttctgatta ttttactgga tatttcaatg gacagtattg gctttggtgg
     2641 atatttcttg tacttggcct gctccttttc ttcagaggat ttgttaatta tctaaaagtc
     2701 agaaacatgt ctgaaagtat ggcagctgct catagaacaa ggtatttctt cttattgtag
     2761 taggcggccg cagcatcgtg actgactgac gatctgcctc gcgcgtttcg gtgatgacgg
     2821 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     2881 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     2941 catgacccag tcacgtagcg atagcggagt gtataattct tgaagacgaa agggcctcgt
     3001 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3061 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3121 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3181 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3241 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3301 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3361 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3421 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3481 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3541 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3601 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3661 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3721 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3781 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3841 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3901 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3961 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4021 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4081 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4141 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4201 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4261 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4321 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     4381 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4441 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4501 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4561 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4621 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4681 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4741 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4801 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4861 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4921 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4981 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     5041 aaattccgac accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga
     5101 agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta
     5161 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     5221 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     5281 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     5341 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     5401 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     5461 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     5521 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     5581 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     5641 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     5701 ct
//
