LOCUS       sequence-430427-        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXB1 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  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 6454)
  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-HOXB1-WT
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     35..54
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     288..305
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    422..443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(458..463,464..481,482..488)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    496..512
                     /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     501..523
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     520..536
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     520..536
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    585..706
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(596..615)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     650..669
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(880..1674)
                     /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(991..1010)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1601..1620
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1741..2070)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1755..1890
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1809..1828)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2055..2075
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2084..2512)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2194..2215)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2406..2425
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2558..2782
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2771..2788
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2808..2826
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2808..2825
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(3449..3889)
                     /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(3951..3970)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3952..3970)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4015..4218)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4044..4064)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4219..4598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4770..4789
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4864..4968
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4969..5037,5038..5829)
                     /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(5187..5206)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6000..6454,1..134)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
       61 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      121 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      181 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      241 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      301 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
      361 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
      421 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
      481 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
      541 attacgccaa gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga
      601 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      661 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga
      721 actccagcat gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc
      781 cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg
      841 gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag
      901 gcgatagaag gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg
      961 gtcagcccat tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg
     1021 atagcggtcc gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc
     1081 caccatgata ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg
     1141 catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc
     1201 cagatcatcc tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg
     1261 tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc
     1321 atcagccatg atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc
     1381 cggcacttcg cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc
     1441 tgcgcaagga acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc
     1501 attcagggca ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag
     1561 ccggaacacg gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag
     1621 cctctccacc caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa
     1681 cgatcctcat cctgtctctt gatcagatcc gaaaatggat atacaagctc ccgggagctt
     1741 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     1801 agaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     1861 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     1921 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     1981 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     2041 tggggacttt ccacacccta actgacacac attccacaga attaattcgc gttaaatttt
     2101 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2161 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2221 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2281 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2341 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2401 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2461 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatac
     2521 cccctagagc cccagctggt tctttccgcc tcagaagcca tagagcccac cgcatcccca
     2581 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     2641 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     2701 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     2761 ggctggcaac tagaaggcac agtcgaggct gatcagcgag ctctagcatt taggtgacac
     2821 tatagaatag ggccctctag accaggcctg gctctgacac cttcgctgtc ttgggtgggt
     2881 ttctcttaac cttcatccag tcgaaggtcc gggccgtggg ggtgttaggt tctgaagggc
     2941 agggtgtttc cttgtcctcg gagaggagat cagcataggc cggtgcaaag ctcgcggtct
     3001 gctcgttccc ataaggggga tgctgcggag gatatggccc cggaccggct ccacctgctc
     3061 cgtagccatc ggacaagccc cctagctggg ccccgtagct cgagggatga aaatagcctc
     3121 cgtctccttc tgattgaccc agagggtagt actgagaagg cccgtagctg gggctgcagg
     3181 cggcaggagc ataccccgag ggcgcggagc tggggaaggg cacccccagg gtcgaaggcg
     3241 gctgctgggc gggatagccg gagttctgct gaaacgcagg gctggacagc cccccaccgt
     3301 agcggccctc gcttgcatag ctgtcaaccg cctgagccga gcttggggga aaggaggttg
     3361 gggcgctgtg ggcgctgtag gcgctgggtc cccggttaca gagtgggtac tctaagaagg
     3421 agttcatcct attatagtcc attatcgccg atacagtcaa ctgtctttga cctttgttac
     3481 tactctcttc cgatgatgat gtcgcactta ttctatgctg tctcaatgtt agaggcatat
     3541 cagtctccac tgaagccaat ctatctgtga cggcatcttt attcacatta tcttgtacaa
     3601 ataatcctgt taacaatgct tttatatcct gtaaagaatc cattttcaaa atcatgtcaa
     3661 ggtcttctcg aggaaaaatc agtagaaata gctgttccag tctttctagc cttgattcca
     3721 cttctgtcag atgtgcccta gtcagcggag accttttggt tttgggagag tagcgacact
     3781 cccagttgtt cttcagacac ttggcgcact tcggtttttc tttggagcac ttgagctttt
     3841 taagtcggca aatatcgcat gcttgttcga tagaagacag tagcttcatg aattccagca
     3901 cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgggtct ccctatagtg
     3961 agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct
     4021 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     4081 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     4141 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     4201 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     4261 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     4321 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     4381 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     4441 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     4501 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     4561 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     4621 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     4681 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     4741 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     4801 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
     4861 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4921 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4981 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5041 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5101 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5161 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5221 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5281 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5341 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5401 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5461 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5521 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5581 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5641 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5701 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacgggaagt
     5761 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5821 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5881 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5941 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6001 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6061 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6421 agaaaggcgg acaggtatcc ggtaagcggc aggg
//
