LOCUS       sequence-443024-        6454 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXB1 AroPERFECT.
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-AroPERFECT
FEATURES             Location/Qualifiers
     source          1..6454
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     548..567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(786..890)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(965..984)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1156..1535
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1536..1739
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1690..1710
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1784..1803
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1784..1802
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1865..2305
                     /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(2928..2946)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2929..2946)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2966..2983)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2972..3196
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3242..3670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3539..3560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3679..3699)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3684..4013
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3864..3999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3926..3945
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4080..4874
                     /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(4134..4153)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4744..4763
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5048..5169
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5085..5104)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5139..5158
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5218..5234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5218..5234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5231..5253)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5242..5258
                     /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(5266..5272,5273..5290,5291..5296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5311..5332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5449..5466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5620..6208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5700..5719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6379..6454,1..716,717..785))
                     /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"
ORIGIN
        1 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
       61 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
      181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
      241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
      361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
      421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
      481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
      541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
      601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
      661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
      721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
      781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      901 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
      961 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     1021 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     1081 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     1141 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     1201 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     1261 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     1321 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     1381 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     1441 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     1501 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     1561 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     1621 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     1681 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     1741 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     1801 gggagaccca agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga
     1861 attcatgaag ctactgtctt ctatcgaaca agcatgcgat atttgccgac ttaaaaagct
     1921 caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg
     1981 ctactctccc aaaaccaaaa ggtctccgct gactagggca catctgacag aagtggaatc
     2041 aaggctagaa agactggaac agctatttct actgattttt cctcgagaag accttgacat
     2101 gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca
     2161 agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc
     2221 tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa
     2281 aggtcaaaga cagttgactg tatcggcgat aatggactat aataggatga actccttaga
     2341 gtacccactc tgtaaccggg gaccctacag cgccagcgcc cacagcgcct ttccaacctc
     2401 ccccccaagc tcgttcgctc aggcggttga cagcgcatat agcgagggcc gcggtggggg
     2461 gtacctgtcc agccctgcgc agcagtttaa ctccggcccc gcccagcagt atccgccttc
     2521 gaccctgggg gtgttccccc ccagctccgc gccctcgtat ggggctcctg ccgcctgcag
     2581 ctaccccagc gggccttctc agccttacct gggtcaatca gaaggagact atggaggcca
     2641 tccctcgagc gggtttgccc agctaggggg cttgtcctac gatggcggag caggtggagc
     2701 ctatggtccg gggccacctc cgcagtatca tccccctggg aacgagcagt acaccgcgag
     2761 cgcaccggcc gcttttgatc tcctctccga ggacaagtac gaaacaccct gcccttcaga
     2821 atatcctaac acccccacgg cccggttcac cgacatgaag gttaagagat ggaacccacc
     2881 caagacagcg aaggtgtcag agccaggcct ggtctagagg gccctattct atagtgtcac
     2941 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     3001 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     3061 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     3121 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3181 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3241 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3841 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3901 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3961 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4021 cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca
     4081 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4141 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4201 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4261 aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4321 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4381 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4441 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4501 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4561 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg
     4621 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4681 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4741 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4801 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4861 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4921 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4981 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5041 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5101 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5161 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6421 tctatttcgt tcatccatag ttgcctgact tccc
//
