LOCUS       sequence-430415-        6279 bp    DNA     circular SYN 10-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXD4 AroPERFECT.
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 10, 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-HOXD4-AroPERFECT
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     374..393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     627..644
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    761..782
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(797..802,803..820,821..827)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    835..851
                     /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     840..862
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     859..875
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     859..875
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    924..1045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(935..954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     989..1008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1219..2013)
                     /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(1330..1349)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1940..1959
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2080..2409)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2094..2229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2148..2167)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2394..2414
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2423..2851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2533..2554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2745..2764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2897..3121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3110..3127
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3147..3165
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3147..3164
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3613..4053)
                     /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(4115..4134)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4116..4134)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4179..4382)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4208..4228)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4383..4762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4934..4953
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5028..5132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5133..5201,5202..5993)
                     /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(5351..5370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6164..6279,1..473)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
       61 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
      121 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
      181 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
      241 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
      301 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      361 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      421 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      481 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      541 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      601 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      661 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      721 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      781 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      841 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag
      901 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
      961 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1021 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     1081 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     1141 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     1201 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     1261 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     1321 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     1381 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     1441 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     1501 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     1561 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     1621 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     1681 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     1741 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     1801 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     1861 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     1921 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     1981 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     2041 atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc
     2101 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc
     2161 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     2221 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     2281 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     2341 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     2401 ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt
     2461 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     2521 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2581 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2641 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2701 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2761 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     2821 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     2881 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     2941 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     3001 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     3061 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     3121 gtcgaggctg atcagcgagc cctagcattt aggtgacact atagaatagg gccctctaga
     3181 cacgactttc ttcattggcc atacaaccgc aggttgtttc agggcatatg tgcctgaagg
     3241 gggttgttta ggataatcgc tttggcttgc ccttgctcca tatgggagag gtgcaggtgg
     3301 gggtgcatag ggtggcgccg gacatggttc cccataaggc gcagcatgtc cgccaggacc
     3361 gtatccgggt tcctgaccat gtcccctgta tgctggaagg gctgatcctg ggccaaaagg
     3421 accactcccg cctggttgtt caaagccatc tggcctgggc aggccaggaa atggttgatc
     3481 cgccccttgt gccccatatc caccatcagc tccttgttcc ccataaagcc caccttgaag
     3541 ttcttcacaa aatggtggtt taggatcaac tttagagtaa ttaaccattg atgacatcac
     3601 catatgtatc gccgatacag tcaactgtct ttgacctttg ttactactct cttccgatga
     3661 tgatgtcgca cttattctat gctgtctcaa tgttagaggc atatcagtct ccactgaagc
     3721 caatctatct gtgacggcat ctttattcac attatcttgt acaaataatc ctgttaacaa
     3781 tgcttttata tcctgtaaag aatccatttt caaaatcatg tcaaggtctt ctcgaggaaa
     3841 aatcagtaga aatagctgtt ccagtctttc tagccttgat tccacttctg tcagatgtgc
     3901 cctagtcagc ggagaccttt tggttttggg agagtagcga cactcccagt tgttcttcag
     3961 acacttggcg cacttcggtt tttctttgga gcacttgagc tttttaagtc ggcaaatatc
     4021 gcatgcttgt tcgatagaag acagtagctt catgaattcc agcacactgg cggccgttac
     4081 tagtggatcc gagctcggta ccaagcttgg gtctccctat agtgagtcgt attaatttcg
     4141 ataagccagt aagcagtggg ttctctagtt agccagagag ctctgcttat atagacctcc
     4201 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     4261 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     4321 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     4381 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     4441 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     4501 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     4561 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     4621 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     4681 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     4741 taactagtca ataatcaatg tcaacgcgta tatctggccc gtacatcgcg aagcagcgca
     4801 aaacgcctaa ccctaagcag attcttcatg caattgtcgg tcaagccttg ccttgttgta
     4861 gcttaaattt tgctcgcgca ctactcagcg acctccaaca cacaagcagg gagcagatac
     4921 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag gggatcggga
     4981 gatctcccga tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5041 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5101 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5161 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5221 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5281 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5341 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5401 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5461 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5521 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5581 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5641 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5701 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5761 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5821 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5881 atggtaagcc ctcccgtatc gtagttatct acacgacggg aagtcaggca actatggatg
     5941 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6001 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6061 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6121 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6181 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6241 cggatcaaga gctaccaact ctttttccga aggtaactg
//
