LOCUS       sequence-392032-        8420 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT human TXNIP with C-terminal 3xFLAG tag in pQCXIP vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8420)
  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 8420)
  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: WT TXNIP-FLAG
FEATURES             Location/Qualifiers
     source          1..8420
                     /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    join(3547..4121,4122..4124,4125..4133)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3714..3731)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3941..3960
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4153..4752
                     /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(4153..4172)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4649..4669
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5126..5551
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5654..5676)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5812..5832)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5817..6146
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5997..6132
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6168..6187
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6268..6285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6439..7027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6519..6538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7198..7989,7990..8058))
                     /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     7821..7840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8059..8163)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8231..8249
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8349..8420,1..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 cctccctgct atatggatgt cattcctgaa gatcaccgat tggagagccc aaccactcct
     3301 ctgctagatg acatggatgg ctctcaagac agccctatct ttatgtatgc ccctgagttc
     3361 aagttcatgc caccaccgac ttatactgag gtggatccct gcatcctcaa caacaatgtg
     3421 cagttaatta acggatcctc aagtttgccg ggcgactata aagaccacga cggtgactac
     3481 aaggaccacg atatagatta caaagatgac gacgataaat gaggaattcc gccccccccc
     3541 cccccccccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     3601 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     3661 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     3721 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     3781 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     3841 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     3901 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
     3961 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     4021 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
     4081 ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccacaagg
     4141 agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     4201 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     4261 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     4321 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     4381 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     4441 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     4501 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     4561 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     4621 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     4681 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     4741 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     4801 catggctccg accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc
     4861 ccaccgactc tagtcgaggg ctgcagcgct gcagaggccg agtgcagaac tgctccaaag
     4921 ggacctcaag gctttccgag ggacactagg ctgactccat cgagccagtg tagagataag
     4981 cttatcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     5041 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     5101 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     5161 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     5221 gatcaaggtc aggaacagat ggaacagggt cgaccctaga gaaccatcag atgtttccag
     5281 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     5341 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     5401 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     5461 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5521 attgactacc cgtcagcggg ggtctttcat ttgggggctc gtccgggatc gggagacccc
     5581 tgcccaggga ccaccgaccc accaccggga ggtaagctgg ctgcctcgcg cgtttcggtg
     5641 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     5701 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     5761 gcgcagccat gacccagtca cgtagcgata gcggagtgta gatccggctg tggaatgtgt
     5821 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5881 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5941 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     6001 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     6061 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     6121 tttttggagg cctaggcttt tgcaaaaagc ttactggctt aactatgcgg catcagagca
     6181 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6241 ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6301 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6361 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6421 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6481 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     6541 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6601 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6661 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6721 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6781 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6841 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     6901 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6961 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7021 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7081 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7141 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7201 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7261 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7321 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7381 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7441 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7501 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7561 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7621 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7681 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7741 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7801 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7861 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7921 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7981 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8041 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8101 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8161 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     8221 aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattag cttggccatt
     8281 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     8341 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     8401 tcatagccca tatatggagt
//
