LOCUS       sequence-212404-        5904 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NFIX2-ERT2 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5904)
  TITLE     mouse NF-1 expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5904)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCMV-NFIX2ERT2
FEATURES             Location/Qualifiers
     source          1..5904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             834..860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2082..3017
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2130..3023
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    3043..3177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3067..3086)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3121..3140
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3288..3304)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3288..3304)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3301..3323)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3312..3328
                     /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)."
     promoter        complement(join(3336..3342,3343..3360,3361..3366))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3381..3402
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3519..3536)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3690..4278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3770..3789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4449..5240,5241..5309))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5072..5091
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5310..5414)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5482..5500
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5538..5560)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5660..5679
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5873..5895
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5887..5904
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5888..5904
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tcgaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg gccgccagcc atgtatcctt
      841 acgatgtacc agactatgct gggcctcacg ggccgatgta tagcccgtac tgcctcaccc
      901 aggatgagtt ccacccgttt atcgaggcgc tgctgcctca cgtccgagcc ttctcctaca
      961 cctggttcaa cctgcaggcg cggaagcgca agtacttcaa gaagcacgag aagcggatgt
     1021 caaaggacga ggagcgcgca gtgaaggacg agctgctggg cgagaagcct gagatcaagc
     1081 agaagtgggc atcccggctg ttggccaagc tgcgcaaaga catccggccc gagttccgcg
     1141 aggactttgt gctaaccatc acgggcaaga agcccccctg ctgcgtgctt tccaaccccg
     1201 accagaaggg caagatccgg cggattgact gcctgcgcca ggctgacaag gtgtggcggc
     1261 tggacctggt catggtgatt ttgtttaaag ggatcccttt ggaaagtact gatggggagc
     1321 ggctctacaa gtcgccccag tgctcgaacc ccggcctgtg tgtccagcca catcacattg
     1381 gagtcacaat caaagaactg gacctttatc tggcttactt tgtccacact ccggaatccg
     1441 gacaatcaga tagttcaaac cagcaaggag atgcggacat caaaccactg cccaacgggc
     1501 acttaagttt ccaggactgc tttgtgacgt ctggggtctg gaatgtgaca gagctggtga
     1561 gagtatcaca gactccagtt gcgactgcat cagggcccaa cttctcactg gcggacctgg
     1621 agagccccag ctactacaac ataaatcaag tgaccctggg aaggcggtcc atcacctccc
     1681 ctccttccac cagcagcacc aagcgcccca agtccatcga cgacagtgag atggagagtc
     1741 cagtagatga tgtgttctat cctgggacag gccgctctcc ggccgctggc agcagccagt
     1801 ctagcggatg gcccaatgac gtggatgcag ggagcccccg ggccacggca tcagcgctgc
     1861 acttcccttc cacgtccatc atccagcagt cgagcccgta tttcacacac ccaaccatcc
     1921 gctaccacca ccaccatggg caggactcgc tgaaggagtt tgtgcagttt gtgtgctctg
     1981 acggctcggg tcaggccacc ggacagcatt cacaacgaca ggcgcctcct ctgccaaccg
     2041 gtttgtcagc atcggatccc gggacggcaa ctttcgggct agctggagac atgagagctg
     2101 ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt
     2161 ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt
     2221 ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc
     2281 tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg
     2341 atttgaccct ccatgatcag gtccaccttc ttgaatgtgc ctggctagag atcctgatga
     2401 ttggtctcgt ctggcgctcc atggagcacc cagtgaagct actgtttgct cctaacttgc
     2461 tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc
     2521 tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca
     2581 aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc
     2641 tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc
     2701 tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc
     2761 tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga
     2821 agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga ggcggcggac gcccaccgcc
     2881 tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg
     2941 ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag
     3001 agggtttccc tgccacagct tgactagtag tagggccgcg gggatccaga catgataaga
     3061 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3121 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3181 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttcg
     3241 gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt
     3301 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3361 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3421 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3481 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3541 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3601 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3661 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3721 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3781 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3841 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3901 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3961 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4021 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4081 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4141 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4201 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4261 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4321 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4381 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4441 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4501 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4561 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4621 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4681 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4741 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4801 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4861 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4921 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4981 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5041 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5101 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5161 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5221 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5281 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5341 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5401 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     5461 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     5521 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     5581 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     5641 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     5701 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     5761 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     5821 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     5881 cgacgttgta aaacgacggc cagt
//
