LOCUS       sequence-422716-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Full length mammalian expression vector for hESR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  TITLE     Vadlamudi lab GFP-ESR plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6477)
  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            "Unpublished"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: GFP-ESR1
FEATURES             Location/Qualifiers
     source          1..6477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             606..1322
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             2190..3125
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2238..3128
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     polyA_signal    3258..3379
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3295..3314)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3349..3368
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3386..3841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3523..3544)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3735..3754
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3868..3972
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3974..4331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3974..4165
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3997..4017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4182..4317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4366..5160
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4366..5160
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4420..4439)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5030..5049
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5348..5367)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5392..5439
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5768..6356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6257..6276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt gagcaagggc gccgagctgt tcaccggcat cgtgcccatc ctgatcgagc
      661 tgaatggcga tgtgaatggc cacaagttca gcgtgagcgg cgagggcgag ggcgatgcca
      721 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgcct gtgccctggc
      781 ccaccctggt gaccaccctg agctacggcg tgcagtgctt ctcacgctac cccgatcaca
      841 tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacatccag gagcgcacca
      901 tcttcttcga ggatgacggc aactacaagt cgcgcgccga ggtgaagttc gagggcgata
      961 ccctggtgaa tcgcatcgag ctgaccggca ccgatttcaa ggaggatggc aacatcctgg
     1021 gcaataagat ggagtacaac tacaacgccc acaatgtgta catcatgacc gacaaggcca
     1081 agaatggcat caaggtgaac ttcaagatcc gccacaacat cgaggatggc agcgtgcagc
     1141 tggccgacca ctaccagcag aataccccca tcggcgatgg ccctgtgctg ctgcccgata
     1201 accactacct gtccacccag agcgccctgt ccaaggaccc caacgagaag cgcgatcaca
     1261 tgatctactt cggcttcgtg accgccgccg ccatcaccca cggcatggat gagctgtaca
     1321 agtccggact cagatctcga gccatgacca tgaccctcca caccaaagca tctgggatgg
     1381 ccctactgca tcagatccaa gggaacgagc tggagcccct gaaccgtccg cagctcaaga
     1441 tccccctgga gcggcccctg ggcgaggtgt acctggacag cagcaagccc gccgtgtaca
     1501 actaccccga gggcgccgcc tacgagttca acgccgcggc cgccgccaac gcgcaggtct
     1561 acggtcagac cggcctcccc tacggccccg ggtctgaggc tgcggcgttc ggctccaacg
     1621 gcctgggggg tttcccccca ctcaacagcg tgtctccgag cccgctgatg ctactgcacc
     1681 cgccgccgca gctgtcgcct ttcctgcagc cccacggcca gcaggtgccc tactacctgg
     1741 agaacgagcc cagcggctac acggtgcgcg aggccggccc gccggcattc tacaggccaa
     1801 attcagataa tcgacgccag ggtggcagag aaagattggc cagtaccaat gacaagggaa
     1861 gtatggctat ggaatctgcc aaggagactc gctactgtgc agtgtgcaat gactatgctt
     1921 caggctacca ttatggagtc tggtcctgtg agggctgcaa ggccttcttc aagagaagta
     1981 ttcaaggaca taacgactat atgtgtccag ccaccaacca gtgcaccatt gataaaaaca
     2041 ggaggaagag ctgccaggcc tgccggctcc gcaaatgcta cgaagtggga atgatgaaag
     2101 gtgggatacg aaaagaccga agaggaggga gaatgttgaa acacaagcgc cagagagatg
     2161 atggggaggg caggggtgaa gtggggtctg ctggagacat gagagctgcc aacctttggc
     2221 caagcccgct catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg
     2281 accagatggt cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc
     2341 ctaccagacc cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg
     2401 agctggttca catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc
     2461 atgatcaggt ccaccttcta gaatgtgcct ggctagagat cctgatgatt ggtctcgtct
     2521 ggcgctccat ggagcaccca gggaagctac tgtttgctcc taacttgctc ttggacagga
     2581 accagggaaa atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat
     2641 ctcggttccg catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt
     2701 tgcttaattc tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg
     2761 accatatcca ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg
     2821 caggcctgac cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc
     2881 acatcaggca catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg
     2941 tggtgcccct ctatgacctg ctgctggaga tgctggacgc ccaccgccta catgcgccca
     3001 ctagccgtgg aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct
     3061 ctacttcatc gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg
     3121 ccacggtcgg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     3181 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     3241 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3301 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3361 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     3421 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     3481 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     3541 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     3601 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     3661 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     3721 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     3781 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     3841 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3901 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3961 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     4021 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4081 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4201 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4261 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4321 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4381 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4441 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4501 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     4561 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4621 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4681 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4741 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4801 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4861 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4921 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4981 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5041 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5101 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5161 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5221 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5281 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     5341 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     5401 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     5461 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     5521 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     5581 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     5641 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5701 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     5761 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5821 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5881 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5941 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6001 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6061 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6121 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6181 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6241 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6301 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6361 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6421 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttat
//
