LOCUS       sequence-443025-        6277 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXD4 AroPLUS PATCHED.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6277)
  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 6277)
  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-HOXD4-AroPLUS_PATCHED
FEATURES             Location/Qualifiers
     source          1..6277
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(14..805,806..874))
                     /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     637..656
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(875..979)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1054..1073)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1245..1624
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1625..1828
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1779..1799
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1873..1892
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1873..1891
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1954..2394
                     /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"
     promoter        complement(2840..2858)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2841..2858)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2878..2895)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2884..3108
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3154..3582
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3241..3260)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3451..3472
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3591..3611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3596..3925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3776..3911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3838..3857
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3992..4786
                     /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(4046..4065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4656..4675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4960..5081
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4997..5016)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5051..5070
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5130..5146)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5130..5146)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5143..5165)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5154..5170
                     /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(5178..5184,5185..5202,5203..5208))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5223..5244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5361..5378)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5532..6120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5612..5631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
       61 ttcgttcatc catagttgcc tgacttcccg tcgtgtagat aactacgata cgggagggct
      121 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
      181 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
      241 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
      301 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
      361 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
      421 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
      481 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      541 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
      601 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
      661 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
      721 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
      781 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
      841 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
      901 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      961 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     1021 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     1081 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     1141 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     1201 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     1261 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     1321 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     1381 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     1441 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     1501 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     1561 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     1621 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     1681 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     1741 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     1801 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     1861 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat
     1921 ccactagtaa cggccgccag tgtgctggaa ttcatgaagc tactgtcttc tatcgaacaa
     1981 gcatgcgata tttgccgact taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc
     2041 aagtgtctga agaacaactg ggagtgtcgc tactctccca aaaccaaaag gtctccgctg
     2101 actagggcac atctgacaga agtggaatca aggctagaaa gactggaaca gctatttcta
     2161 ctgatttttc ctcgagaaga ccttgacatg attttgaaaa tggattcttt acaggatata
     2221 aaagcattgt taacaggatt atttgtacaa gataatgtga ataaagatgc cgtcacagat
     2281 agattggctt cagtggagac tgatatgcct ctaacattga gacagcatag aataagtgcg
     2341 acatcatcat cggaagagag tagtaacaaa ggtcaaagac agttgactgt atcggcgatc
     2401 atggttatga gcagttacat ggtcaatagc aaatacgtcg atcctaaatt ttatccacct
     2461 tgtgaagaat atctccaagg cggatattac cttggagaac aaggagcaga ttattatggt
     2521 ggcggagccc aaggagcgga ttttcaacca cccggactgt attaccctcg accagatttt
     2581 ggcgaacaac cattttatgg tgggtccggt cctggcggta gcgctctccc cgctaggcat
     2641 gggcaggaac ctgggccagg cggacattac tatgctgccc ctggtgaacc ttgtcctccg
     2701 cccgccccac caccacctct tccaggggct agggcatatt attcacaatc agatccgaaa
     2761 cccccatctg gcaccgccaa acaaccagcg gtagtgtact atccttggat gaagaaagta
     2821 tgagtctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     2881 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     2941 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     3001 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     3061 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     3121 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3181 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3241 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3301 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3361 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3421 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3481 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3541 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3841 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3901 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3961 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4141 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4501 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4621 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4801 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4861 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4921 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4981 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5041 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5101 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5161 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5221 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5281 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5341 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5401 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5461 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5521 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5581 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5641 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5701 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5761 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5821 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5881 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5941 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6001 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6061 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6121 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6181 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6241 tgaagtttta aatcaatcta aagtatatat gagtaaa
//
