LOCUS       sequence-427389-        7202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-ERalpha in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7202)
  AUTHORS   Rona G, Miwatani-Minter B, Zhang Q, Goldberg HV, Kerzhnerman MA, 
            Howard JB, Simoneschi D, Lane E, Hobbs JW, Sassani E, Wang AA, 
            Keegan S, Laverty DJ, Piett CG, Pongor LS, Xu ML, Andrade J, Thomas 
            A, Sicinski P, Askenazi M, Ueberheide B, Fenyo D, Nagel ZD, Pagano M
  TITLE     CDK-independent role of D-type cyclins in regulating DNA mismatch 
            repair.
  JOURNAL   Mol Cell. 2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010.
  PUBMED    38458201
REFERENCE   2  (bases 1 to 7202)
  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: "Mol Cell. 
            2024 Feb 29:S1097-2765(24)00129-1. doi: 
            10.1016/j.molcel.2024.02.010."
COMMENT     Sequence Label: HA-ERalpha
FEATURES             Location/Qualifiers
     source          1..7202
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1786..2721
                     /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             1834..2727
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     promoter        complement(2755..2773)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2756..2773)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2793..2810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2799..3023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3069..3497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3156..3175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3366..3387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3506..3526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3511..3840
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3691..3826
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3753..3772
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3907..4701
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3961..3980)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4571..4590
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4875..4996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4912..4931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4966..4985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5045..5061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5045..5061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5058..5080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5069..5085
                     /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(5093..5099,5100..5117,5118..5123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5276..5293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5447..6035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5527..5546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6206..6997,6998..7066))
                     /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     6829..6848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7067..7171)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gcctacccct acgacgtgcc cgactacgcc tccctcggat ccaccatgac cctccacacc
      961 aaagcatctg ggatggccct actgcatcag atccaaggga acgagctgga gcccctgaac
     1021 cgtccgcagc tcaagatccc cctggagcgg cccctgggcg aggtgtacct ggacagcagc
     1081 aagcccgccg tgtacaacta ccccgagggc gccgcctacg agttcaacgc cgcggccgcc
     1141 gccaacgcgc aggtctacgg tcagaccggc ctcccctacg gccccgggtc tgaggctgcg
     1201 gcgttcggct ccaacggcct ggggggtttc cccccactca acagcgtgtc tccgagcccg
     1261 ctgatgctac tgcacccgcc gccgcagctg tcgcctttcc tgcagcccca cggccagcag
     1321 gtgccctact acctggagaa cgagcccagc ggctacacgg tgcgcgaggc cggcccgccg
     1381 gcattctaca ggccaaattc agataatcga cgccagggtg gcagagaaag attggccagt
     1441 accaatgaca agggaagtat ggctatggaa tctgccaagg agactcgcta ctgtgcagtg
     1501 tgcaatgact atgcttcagg ctaccattat ggagtctggt cctgtgaggg ctgcaaggcc
     1561 ttcttcaaga gaagtattca aggacataac gactatatgt gtccagccac caaccagtgc
     1621 accattgata aaaacaggag gaagagctgc caggcctgcc ggctccgcaa atgctacgaa
     1681 gtgggaatga tgaaaggtgg gatacgaaaa gaccgaagag gagggagaat gttgaaacac
     1741 aagcgccaga gagatgatgg ggagggcagg ggtgaagtgg ggtctgctgg agacatgaga
     1801 gctgccaacc tttggccaag cccgctcatg atcaaacgct ctaagaagaa cagcctggcc
     1861 ttgtccctga cggccgacca gatggtcagt gccttgttgg atgctgagcc ccccatactc
     1921 tattccgagt atgatcctac cagacccttc agtgaagctt cgatgatggg cttactgacc
     1981 aacctggcag acagggagct ggttcacatg atcaactggg cgaagagggt gccaggcttt
     2041 gtggatttga ccctccatga tcaggtccac cttctagaat gtgcctggct agagatcctg
     2101 atgattggtc tcgtctggcg ctccatggag cacccaggga agctactgtt tgctcctaac
     2161 ttgctcttgg acaggaacca gggaaaatgt gtagagggca tggtggagat cttcgacatg
     2221 ctgctggcta catcatctcg gttccgcatg atgaatctgc agggagagga gtttgtgtgc
     2281 ctcaaatcta ttattttgct taattctgga gtgtacacat ttctgtccag caccctgaag
     2341 tctctggaag agaaggacca tatccaccga gtcctggaca agatcacaga cactttgatc
     2401 cacctgatgg ccaaggcagg cctgaccctg cagcagcagc accagcggct ggcccagctc
     2461 ctcctcatcc tctcccacat caggcacatg agtaacaaag gcatggagca tctgtacagc
     2521 atgaagtgca agaacgtggt gcccctctat gacctgctgc tggagatgct ggacgcccac
     2581 cgcctacatg cgcccactag ccgtggaggg gcatccgtgg aggagacgga ccaaagccac
     2641 ttggccactg cgggctctac ttcatcgcat tccttgcaaa agtattacat cacgggggag
     2701 gcagagggtt tccctgccac ggtctgactc gagcatgcat ctagagggcc ctattctata
     2761 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2821 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2881 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2941 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3001 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3061 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     3421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     3481 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     3541 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     3601 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3661 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     3721 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     3781 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     3841 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     3901 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3961 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4021 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4081 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4141 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4201 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4261 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4321 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4381 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     4441 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4501 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4561 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4621 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4681 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4741 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4801 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4861 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4921 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4981 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5041 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5101 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5161 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5221 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5281 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5341 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5401 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5461 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5521 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5581 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5641 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5701 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5761 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5821 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5881 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5941 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6001 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6061 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6121 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6181 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6241 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6301 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6361 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6421 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6481 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6541 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6601 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6661 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6721 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6781 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6841 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6901 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6961 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7021 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7081 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7141 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7201 tc
//
