LOCUS       sequence-214845-        5998 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of NFIB2-ERT2 fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5998)
  TITLE     mouse NF-1 expression vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5998)
  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            "Unpublished"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-NFIB2ERt2
FEATURES             Location/Qualifiers
     source          1..5998
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..323
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        324..527
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     478..498
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     524..548
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          665..761
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             827..853
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2132..3067
                     /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             2180..3073
                     /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    3130..3264
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3154..3173)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3208..3227
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3375..3391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3375..3391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3388..3410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3399..3415
                     /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(3423..3429,3430..3447,3448..3453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3468..3489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3606..3623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3777..4365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3857..3876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4536..5327,5328..5396))
                     /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     5159..5178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5397..5501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5569..5587
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5625..5647)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5747..5766
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5960..5982
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5974..5991
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5975..5991
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 agcttgcatg cctgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
       61 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      121 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      181 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      241 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      301 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      361 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      421 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      481 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      541 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
      601 gatccagcct ccggactcta gaggatccgg tactcgagga actgaaaaac cagaaagtta
      661 actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg gtggtgcaaa
      721 tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg gaagtgttac
      781 ttctgctcta aaagctgcgg aattgtaccc gcggccgcca gccatgtatc cttacgatgt
      841 accagactat gctgggcctc acgggccgat gatgtattct cccatctgtc tcactcagga
      901 tgaattccac ccatttattg aggcacttct tcctcacgtc cgtgcaattg cctatacttg
      961 gttcaacctg caggctcgaa aacgcaagta ctttaaaaag catgagaaac gaatgtcgaa
     1021 ggatgaagaa agggcagtca aagacgagct gctcagtgag aagcccgaaa tcaagcagaa
     1081 gtgggcatcc aggctcctgg ccaaactgcg caaagatatc cgccaggagt accgggagga
     1141 ctttgtgctt accgtgactg gcaagaagca cccgtgctgt gtcttatcca atccagacca
     1201 gaagggtaag attaggagga tcgactgcct gcgacaggca gacaaagtct ggcgtctgga
     1261 tctagtcatg gtgatcctgt tcaaaggcat ccctttggag agtacggatg gagagcgact
     1321 catgaagtcc ccgcactgca caaacccagc actttgtgtt cagccacacc acatcacagt
     1381 atcagttaag gagcttgact tgtttttggc atactacgtg caggagcaag attctggaca
     1441 atcaggaagt ccaagccaca gtgatcctgc caagaatcct ccagggtacc tcgaggacag
     1501 ctttgtaaaa tccggagtct tcaatgtatc agagcttgtg agagtatcca gaacacccat
     1561 aacccaggga actggagtca acttcccaat cggagaaatt cccagccaac catactatca
     1621 tgacatgaac tctggtgtga acctgcagag gtcgctgtct tctccaccga gcagcaaaag
     1681 acccaaaact atatctatag atgaaaatat ggagccaagt cctacaggag acttttaccc
     1741 ctctccaaat tcaccagctg ctggaagtcg aacatggcat gaacgagatc aagatatgtc
     1801 ttctccaact acaatgaaga agcctgagaa gccactgttt agctctacat ctccacagga
     1861 ttcttcccca agattgagca ctttccccca gcaccatcat cccggaatac ctggagtcgc
     1921 gcacagtgtc atctcaactc gaactccacc tccgccctca ccgttgccat ttccgacgca
     1981 agctatcctt cctccggcac cttccagcta cttctctcat ccaacaatca gatatcctcc
     2041 tcacctgaat cctcaggata ctctgaagaa ctacgtacct tcttatgacc catccagtcc
     2101 tcaaacgagc cagtcctggt acctggggct agctggagac atgagagctg ccaacctttg
     2161 gccaagcccg ctcatgatca aacgctctaa gaagaacagc ctggccttgt ccctgacggc
     2221 cgaccagatg gtcagtgcct tgttggatgc tgagcccccc atactctatt ccgagtatga
     2281 tcctaccaga cccttcagtg aagcttcgat gatgggctta ctgaccaacc tggcagacag
     2341 ggagctggtt cacatgatca actgggcgaa gagggtgcca ggctttgtgg atttgaccct
     2401 ccatgatcag gtccaccttc ttgaatgtgc ctggctagag atcctgatga ttggtctcgt
     2461 ctggcgctcc atggagcacc cagtgaagct actgtttgct cctaacttgc tcttggacag
     2521 gaaccaggga aaatgtgtag agggcatggt ggagatcttc gacatgctgc tggctacatc
     2581 atctcggttc cgcatgatga atctgcaggg agaggagttt gtgtgcctca aatctattat
     2641 tttgcttaat tctggagtgt acacatttct gtccagcacc ctgaagtctc tggaagagaa
     2701 ggaccatatc caccgagtcc tggacaagat cacagacact ttgatccacc tgatggccaa
     2761 ggcaggcctg accctgcagc agcagcacca gcggctggcc cagctcctcc tcatcctctc
     2821 ccacatcagg cacatgagta acaaaggcat ggagcatctg tacagcatga agtgcaagaa
     2881 cgtggtgccc ctctatgacc tgctgctgga ggcggcggac gcccaccgcc tacatgcgcc
     2941 cactagccgt ggaggggcat ccgtggagga gacggaccaa agccacttgg ccactgcggg
     3001 ctctacttca tcgcattcct tgcaaaagta ttacatcacg ggggaggcag agggtttccc
     3061 tgccacagct tgagtagtag aagggcgaat tccagcacac tggcggccgt tactagctag
     3121 ggccgcgggg atccagacat gataagatac attgatgagt ttggacaaac cacaactaga
     3181 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     3241 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     3301 cagggggagg tgtgggaggt tttttcggat cctctagagt cgacctgcag gcatgcaagc
     3361 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5521 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5581 acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     5641 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     5701 ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag
     5761 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5821 taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     5881 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     5941 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattcg
//
