LOCUS       sequence-210948-        6475 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human TFAP4 (E135A/S139A) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6475)
  AUTHORS   D'Annibale S, Kim J, Magliozzi R, Low TY, Mohammed S, Heck AJ, 
            Guardavaccaro D
  TITLE     Proteasome-dependent degradation of transcription factor activating 
            enhancer-binding protein 4 (TFAP4) controls mitotic division.
  JOURNAL   J Biol Chem. 2014 Mar 14;289(11):7730-7. doi: 
            10.1074/jbc.M114.549535. Epub 2014 Feb 5.
  PUBMED    24500709
REFERENCE   2  (bases 1 to 6475)
  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"; doi: 
            "10.1074/jbc.M114.549535"; journalName: "J Biol Chem"; date: 
            "2014-03-14- 14"; volume: "289"; issue: "11"; pages: "7730-7"
COMMENT     Sequence Label: pcDNA3-HA-TFAP4(E135A/S139A)
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(67..86)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        258..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        638..841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     886..905
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        886..904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             976..1002
                     /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             976..1002
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2051..2069)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2052..2069)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2089..2106)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2095..2319
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2365..2793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2452..2471)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2662..2683
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2802..2822)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2807..3136
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2987..3122
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3049..3068
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3203..3997
                     /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(3257..3276)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3867..3886
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4171..4292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4208..4227)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4262..4281
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4341..4357)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4341..4357)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4354..4376)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4365..4381
                     /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(4389..4395,4396..4413,4414..4419))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4434..4455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4572..4589)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4743..5331)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4823..4842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5502..6293,6294..6362))
                     /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     6125..6144
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6363..6467)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
       61 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
      121 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
      181 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
      241 ggccagatat acgcgttgac attgattatt gactagttat taatagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      841 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
      901 tagggagacc caagcttggt accgagctcg gatccactag taacggccgc cagtgtgctg
      961 gaattcgcca ccatgtaccc ctacgacgtg cccgactacg ccgagtattt catggtgccc
     1021 actcagaagg tgccctcttt gcaacatttc aggaaaacag agaaagaagt gataggaggg
     1081 ctctgtagcc ttgccaacat tccactaacc cccgagactc agcgggacca ggagcggcgg
     1141 attcggcggg agatcgccaa cagcaacgag cggagacgca tgcagagcat caacgcggga
     1201 ttccagtccc tcaagaccct catcccccac acagacggag agaagctcag caaggcagcc
     1261 attctccagc agacagccga gtacatcttc tccctggagc aggagaagac caggctcttg
     1321 cagcagaaca cacagctcaa gcgcttcatc caggagctga gcggctcgtc ccccaagcga
     1381 cggcgggcag aggacaagga cgcaggcata ggcgccccgg acatctggga ggacgagaag
     1441 gcggaggacc tgcggcggga gatgattgag ctgcggcagc agctggacaa ggagcgctcg
     1501 gtgcgcatga tgctggagga gcaggtgcgc tcgctggagg cccacatgta cccggaaaag
     1561 ctcaaggtga ttgcgcagca ggtgcagctg cagcagcagc aggaacaggt gaggctgctg
     1621 caccaggaga agctggagcg ggaacagcag cagctgcgga cccagcttct gccccctccg
     1681 gcccccaccc accaccccac ggtgatcgtg ccagcaccgc ctcctcctcc ctcccaccac
     1741 atcaatgtcg tcaccatggg cccctcctcg gtcatcaact ctgtttccac atcccggcaa
     1801 aatctggaca ccatcgtgca ggcaatccag cacatcgagg gcacccagga aaagcaggag
     1861 ctggaggagg agcagcggcg agctgtcatc gtgaagcctg tccgcagctg cccggaggcc
     1921 cccacctctg acaccgcctc cgactccgag gcctcagaca gtgacgccat ggaccagagc
     1981 cgggaggagc cgtcggggga cggggagctt ccctgagcgg ccgctcgagc atgcatctag
     2041 agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc
     2101 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2161 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2221 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2281 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2341 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2581 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2701 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2761 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2821 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2881 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2941 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3001 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3061 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3121 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3181 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3241 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3301 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3361 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3421 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3481 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3541 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3601 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3661 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3721 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3781 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3841 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3901 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3961 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4021 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4081 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4141 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4201 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4261 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4321 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4381 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4441 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4501 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4561 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4621 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4681 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4741 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4861 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4921 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4981 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5041 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5101 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5161 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5221 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5281 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5341 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5401 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5461 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5521 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5581 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5641 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5701 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5761 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5821 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5881 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5941 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     6001 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6061 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6121 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6181 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6241 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6301 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6361 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6421 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttc
//
