LOCUS       sequence-392107-        8389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TXNIP py motif mutant with C-terminal 3xFLAG tag in pQCXIP vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8389)
  AUTHORS   Qualls-Histed SJ, Nielsen CP, MacGurn JA
  TITLE     Lysosomal trafficking of the glucose transporter GLUT1 requires 
            sequential regulation by TXNIP and ubiquitin.
  JOURNAL   iScience. 2023 Feb 6;26(3):106150. doi: 10.1016/j.isci.2023.106150. 
            eCollection 2023 Mar 17.
  PUBMED    36890792
REFERENCE   2  (bases 1 to 8389)
  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: "iScience.";
            date: "2023-02-6"; volume: "26(3):106150. doi"; pages: " 
            10.1016/j.isci.2023.106150. eCollection 2023 Mar 17"
COMMENT     Sequence Label: TXNIP-py-FLAG
FEATURES             Location/Qualifiers
     source          1..8389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             3454..3519
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3627..4089
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3683..3700)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3910..3929
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4122..4721
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4122..4141)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4618..4638
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5095..5520
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5623..5645)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5781..5801)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5786..6115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5966..6101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6137..6156
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6237..6254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6408..6996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6488..6507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7167..7958,7959..8027))
                     /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     7790..7809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8028..8132)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8200..8218
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8318..8389,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     2221 cggacctgca ggaattgatc cgcggccgca atggtgatgt tcaagaagat caagtctttt
     2281 gaggtggtct ttaacgaccc tgaaaaggtg tacggcagtg gcgagaaggt ggctggccgg
     2341 gtgatagtgg aggtgtgtga agttactcgt gtcaaagccg ttaggatcct ggcttgcgga
     2401 gtggctaaag tgctttggat gcagggatcc cagcagtgca aacagacttc ggagtacctg
     2461 cgctatgaag acacgcttct tctggaagac cagccaacag gtgagaatga gatggtgatc
     2521 atgagacctg gaaacaaata tgagtacaag ttcggctttg agcttcctca ggggcctctg
     2581 ggaacatcct tcaaaggaaa atatgggtgt gtagactact gggtgaaggc ttttcttgac
     2641 cgcccgagcc agccaactca agagacaaag aaaaactttg aagtagtgga tctggtggat
     2701 gtcaataccc ctgatttaat ggcacctgtg tctgctaaaa aagaaaagaa agtttcctgc
     2761 atgttcattc ctgatgggcg ggtgtctgtc tctgctcgaa ttgacagaaa aggattctgt
     2821 gaaggtgatg agatttccat ccatgctgac tttgagaata catgttcccg aattgtggtc
     2881 cccaaagctg ccattgtggc ccgccacact taccttgcca atggccagac caaggtgctg
     2941 actcagaagt tgtcatcagt cagaggcaat catattatct cagggacatg cgcatcatgg
     3001 cgtggcaaga gccttcgggt tcagaagatc aggccttcta tcctgggctg caacatcctt
     3061 cgagttgaat attccttact gatctatgtt agcgttcctg gatccaagaa ggtcatcctt
     3121 gacctgcccc tggtaattgg cagcagatca ggtctaagca gcagaacatc cagcatggcc
     3181 agccgaacca gctctgagat gagttgggta gatctgaaca tccctgatac cccagaagct
     3241 cctgcctgcg ccatggatgt cattcctgaa gatcaccgat tggagagccc aaccactcct
     3301 ctgctagatg acatggatgg ctctcaagac agccctatct ttatgtatgc ccctgagttc
     3361 aagttcatgc caccagccac tgccactgag gtggatccct gcatcctcaa caacaatgtg
     3421 cagttaatta acggatcctc aagtttgccg ggcgactata aagaccacga cggtgactac
     3481 aaggaccacg atatagatta caaagatgac gacgataaat gnnnntcccc cccccctaac
     3541 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     3601 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     3661 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     3721 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     3781 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     3841 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3901 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3961 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4021 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     4081 cgatgataag cttgccacaa cccacaagga gacgaccttc catgaccgag tacaagccca
     4141 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     4201 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     4261 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     4321 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     4381 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     4441 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     4501 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     4561 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     4621 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     4681 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc cacgacccgc
     4741 agcgcccgac cgaaaggagc gcacgacccc atggctccga ccgaagccac ccggggcggc
     4801 cccgccgacc ccgcacccgc ccccgaggcc caccgactct agtcgagggc tgcagcgctg
     4861 cagaggccga gtgcagaact gctccaaagg gacctcaagg ctttccgagg gacactaggc
     4921 tgactccatc gagccagtgt agagataagc ttatcgatta gtccaatttg ttaaagacag
     4981 gatatcagtg gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga
     5041 gtacgagcca tagataaaat aaaagatttt atttagtctc cagaaaaagg ggggaatgaa
     5101 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
     5161 aaatacataa ctgagaatag agaagttcag atcaaggtca ggaacagatg gaacagggtc
     5221 gaccctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     5281 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     5341 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     5401 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5461 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     5521 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     5581 gtaagctggc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     5641 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     5701 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     5761 cggagtgtag atccggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     5821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     5881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     5941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     6001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6121 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     6181 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     6241 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6301 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6361 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6421 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6481 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6541 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6601 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6661 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6721 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6781 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6841 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6901 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6961 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7021 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7081 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7141 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7201 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7261 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7321 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7381 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7441 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca
     7501 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7561 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7621 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7681 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7741 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     7801 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7861 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7921 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7981 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8041 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8101 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8161 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8221 tttcgtcttc aagaattagc ttggccattg catacgttgt atccatatca taatatgtac
     8281 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     8341 taatagtaat caattacggg gtcattagtt catagcccat atatggagt
//
