LOCUS       sequence-430421-        6279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXC4 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6279)
  AUTHORS   Naderi J, Magalhaes AP, Kibar G, Stik G, Zhang Y, Mackowiak SD, 
            Wieler HM, Rossi F, Buschow R, Christou-Kent M, Alcoverro-Bertran M,
            Graf T, Vingron M, Hnisz D
  TITLE     An activity-specificity trade-off encoded in human transcription 
            factors.
  JOURNAL   Nat Cell Biol. 2024 Aug;26(8):1309-1321. doi: 
            10.1038/s41556-024-01411-0. Epub 2024 Jul 5.
  PUBMED    38969762
REFERENCE   2  (bases 1 to 6279)
  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: 
            "10.1038/s41556-024-01411-0"; journalName: "Nat Cell Biol"; date: 
            "2024-08"; volume: "26"; issue: "8"; pages: "1309-1321"
COMMENT     Sequence Label: pGAL4-DBD-HOXC4-WT
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     165..184
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     418..435
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    552..573
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(588..593,594..611,612..618)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    626..642
                     /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)."
     primer_bind     631..653
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     650..666
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     650..666
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    715..836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(726..745)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     780..799
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1010..1804)
                     /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(1121..1140)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1731..1750
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1871..2200)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1885..2020
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1939..1958)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2185..2205
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2214..2642)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2324..2345)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2536..2555
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2688..2912
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2901..2918
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2938..2956
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2938..2955
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3404..3844)
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     primer_bind     complement(3906..3925)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3907..3925)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3970..4173)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3999..4019)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4174..4553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4725..4744
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4819..4923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4924..4992,4993..5784)
                     /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     complement(5142..5161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5955..6279,1..264)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
       61 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      121 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      181 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      241 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      301 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
      361 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      421 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
      481 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
      541 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
      601 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
      661 tatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat
      721 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
      781 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg
      841 tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg
      901 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat
      961 ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact
     1021 cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca
     1081 cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg
     1141 ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc
     1201 ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct
     1261 cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat
     1321 gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct
     1381 cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc
     1441 gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga
     1501 gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt
     1561 cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt
     1621 cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct
     1681 gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat
     1741 agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa
     1801 tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc gaaaatggat atacaagctc
     1861 ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata
     1921 gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg
     1981 gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga
     2041 ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     2101 gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     2161 ctggggagcc tggggacttt ccacacccta actgacacac attccacaga attaattcgc
     2221 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2281 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2341 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2401 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2461 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2521 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2581 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2641 gtggggatac cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac
     2701 cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac
     2761 cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta
     2821 ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca
     2881 aacaacagat ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctagcatt
     2941 taggtgacac tatagaatag ggccctctag acattttcat ccatgggtag actatgggtt
     3001 gcttgctggc ggcgctggag ggatggtcgg gggctggctg gctgcaggct ggcggggctg
     3061 gggacgggga ggcggaggcg cctgagagag gcgccggctc gcagagcgac tgtgatttct
     3121 cggggtggtg gtggcccgcc tgggccggcc cgtggcctcg cgaattgccg ggcccctgga
     3181 gactggtgca gctatactgg cgctcagggt agctagggcg cggaggcggt ggtgggtaca
     3241 gctcctggtg gtgatgctgg aatcccgatt ccctggtccg gccgtaatat tccggactgt
     3301 gttcagggat gtagctattt tgcgaatatt cttcgcatgg aggaaatttc ggatcgatgt
     3361 agttagagtc catcaaatac gagctcatga tcatatgtat cgccgataca gtcaactgtc
     3421 tttgaccttt gttactactc tcttccgatg atgatgtcgc acttattcta tgctgtctca
     3481 atgttagagg catatcagtc tccactgaag ccaatctatc tgtgacggca tctttattca
     3541 cattatcttg tacaaataat cctgttaaca atgcttttat atcctgtaaa gaatccattt
     3601 tcaaaatcat gtcaaggtct tctcgaggaa aaatcagtag aaatagctgt tccagtcttt
     3661 ctagccttga ttccacttct gtcagatgtg ccctagtcag cggagacctt ttggttttgg
     3721 gagagtagcg acactcccag ttgttcttca gacacttggc gcacttcggt ttttctttgg
     3781 agcacttgag ctttttaagt cggcaaatat cgcatgcttg ttcgatagaa gacagtagct
     3841 tcatgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg
     3901 ggtctcccta tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt
     3961 tagccagaga gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     4021 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     4081 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     4141 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     4201 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     4261 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     4321 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     4381 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     4441 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg
     4501 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     4561 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     4621 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     4681 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     4741 tgtactgaga gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc
     4801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4861 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4921 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4981 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5041 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5101 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5161 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5221 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5281 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5341 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5401 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5461 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5521 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5581 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5641 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5701 tacacgacgg gaagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5761 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5821 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5881 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5941 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6001 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6061 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6121 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6181 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6241 tagttaccgg ataaggcgca gcggtcgggc tgaacgggg
//
