LOCUS       sequence-437910-        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"
     CDS             complement(join(40..831,832..900))
                     /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     663..682
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(901..1005)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1080..1099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1271..1650
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1651..1854
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1805..1825
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1899..1918
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1899..1917
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1980..2420
                     /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(3043..3061)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3044..3061)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3081..3098)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3087..3311
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3357..3785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3444..3463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3654..3675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3794..3814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3799..4128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4041..4060
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4195..4989
                     /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(4249..4268)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4859..4878
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5163..5284
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5200..5219)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5254..5273
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5333..5349)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5333..5349)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5346..5368)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5357..5373
                     /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(5381..5387,5388..5405,5406..5411))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5426..5447
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5564..5581)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5735..6323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5815..5834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
       61 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac ttcccgtcgt
      121 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      181 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      241 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
      301 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
      361 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
      421 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
      481 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
      541 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
      601 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
      661 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
      721 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
      781 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
      841 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
      901 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
      961 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1021 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     1081 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1141 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1201 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1261 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     1321 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1381 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1441 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1501 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1561 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1621 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     1681 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1741 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1801 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     1861 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     1921 acccaagctt ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca
     1981 tgaagctact gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt
     2041 gctccaaaga aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact
     2101 ctcccaaaac caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc
     2161 tagaaagact ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt
     2221 tgaaaatgga ttctttacag gatataaaag cattgttaac aggattattt gtacaagata
     2281 atgtgaataa agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa
     2341 cattgagaca gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc
     2401 aaagacagtt gactgtatcg gcgataatgg actataatag gatgaactcc ttcttagagt
     2461 acccactctg taaccgggga cccagcgcct acagcgccca cagcgcccca acctcctttc
     2521 ccccaagctc ggctcaggcg gttgacagct atgcaagcga gggccgctac ggtggggggc
     2581 tgtccagccc tgcgtttcag cagaactccg gctatcccgc ccagcagccg ccttcgaccc
     2641 tgggggtgcc cttccccagc tccgcgccct cggggtatgc tcctgccgcc tgcagcccca
     2701 gctacgggcc ttctcagtac taccctctgg gtcaatcaga aggagacgga ggctattttc
     2761 atccctcgag ctacggggcc cagctagggg gcttgtccga tggctacgga gcaggtggag
     2821 ccggtccggg gccatatcct ccgcagcatc ccccttatgg gaacgagcag accgcgagct
     2881 ttgcaccggc ctatgctgat ctcctctccg aggacaagga aacaccctgc ccttcagaac
     2941 ctaacacccc cacggcccgg accttcgact ggatgaaggt taagagaaac ccacccaaga
     3001 cagcgaaggt gtcagagcca ggcctggtct agagggccct attctatagt gtcacctaaa
     3061 tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3121 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3181 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3241 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3301 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3361 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3421 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3481 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3541 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3601 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4141 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4201 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4261 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4321 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4381 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4441 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4501 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4561 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4621 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4681 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4741 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4801 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4861 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4921 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4981 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5041 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5101 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5161 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5221 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5281 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5341 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5401 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5461 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5521 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5581 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5641 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6241 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6301 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6361 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6421 cctagatcct tttaaattaa aaatgaagtt ttaa
//
