LOCUS       sequence-430423-        6280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXC4 AroPERFECT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6280)
  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 6280)
  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-HOXC4-AroPERFECT
FEATURES             Location/Qualifiers
     source          1..6280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    116..340
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     329..346
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        366..384
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     366..383
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(833..1273)
                     /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(1335..1354)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1336..1354)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1399..1602)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1428..1448)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1603..1982
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2154..2173
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2248..2352
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2353..2421,2422..3213)
                     /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(2571..2590)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3384..3972
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3873..3892
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4126..4143
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4260..4281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4296..4301,4302..4319,4320..4326)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4334..4350
                     /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     4339..4361
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4358..4374
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4358..4374
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4423..4544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4434..4453)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4488..4507
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4718..5512)
                     /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(4829..4848)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5439..5458
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5579..5908)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5593..5728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5647..5666)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5893..5913
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(5922..6280,1..70))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6032..6053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6244..6263
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
       61 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
      121 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
      181 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
      241 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
      301 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgagct
      361 ctagcattta ggtgacacta tagaataggg ccctctagac gattttccac attgggacta
      421 tgggttgctt gctggcgtag gcgctggagg gatggtcggg ggctggctga tagctgcagg
      481 ctggcggggc tggggacggg gagtaggcgg aggcgcctga gagaggcgcc ggctcgtagc
      541 agagcgactg tgatttctcg gggtggtgat agtggcccgc ctgggccggc ccgtggcctc
      601 ggtacgaatt gccgggcccc tggagactgg tgcaatagct ctggcgctca gggctagggc
      661 gcggagggta cggtggtggc agctcctggt ggtgatgctg gaatcccgat tccctggtcc
      721 ggccttccgg actgtagtgt tcagggatgc tattttgcga ttcttcaaag catggaggtt
      781 tcggatcgat gttagagtca tacatcaacg agctcatgat catatgtatc gccgatacag
      841 tcaactgtct ttgacctttg ttactactct cttccgatga tgatgtcgca cttattctat
      901 gctgtctcaa tgttagaggc atatcagtct ccactgaagc caatctatct gtgacggcat
      961 ctttattcac attatcttgt acaaataatc ctgttaacaa tgcttttata tcctgtaaag
     1021 aatccatttt caaaatcatg tcaaggtctt ctcgaggaaa aatcagtaga aatagctgtt
     1081 ccagtctttc tagccttgat tccacttctg tcagatgtgc cctagtcagc ggagaccttt
     1141 tggttttggg agagtagcga cactcccagt tgttcttcag acacttggcg cacttcggtt
     1201 tttctttgga gcacttgagc tttttaagtc ggcaaatatc gcatgcttgt tcgatagaag
     1261 acagtagctt catgaattcc agcacactgg cggccgttac tagtggatcc gagctcggta
     1321 ccaagcttgg gtctccctat agtgagtcgt attaatttcg ataagccagt aagcagtggg
     1381 ttctctagtt agccagagag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     1441 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     1501 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     1561 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     1621 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     1681 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     1741 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     1801 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     1861 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     1921 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     1981 tcaacgcgta tatctggccc gtacatcgcg aagcagcgca aaacgcctaa ccctaagcag
     2041 attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca
     2101 ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc
     2161 agagcagatt gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg
     2221 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2281 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2341 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2401 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2461 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2521 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2581 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2641 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2701 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2761 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2821 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2881 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2941 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3001 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3061 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3121 gtagttatct acacgacggg aagtcaggca actatggatg aacgaaatag acagatcgct
     3181 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3241 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3301 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3361 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3421 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3481 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3541 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3601 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3661 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3721 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3781 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3841 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3901 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3961 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     4021 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     4081 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     4141 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     4201 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     4261 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     4321 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     4381 tacgccaagc tctagctaga ggtcgacggt atacagacat gataagatac attgatgagt
     4441 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     4501 ctattgcttt atttgtaacc attataagct gcaataaaca agttggggtg ggcgaagaac
     4561 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     4621 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     4681 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     4741 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     4801 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     4861 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     4921 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     4981 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     5041 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     5101 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     5161 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     5221 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     5281 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     5341 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     5401 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     5461 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     5521 atcctcatcc tgtctcttga tcagatccga aaatggatat acaagctccc gggagctttt
     5581 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag
     5641 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
     5701 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
     5761 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     5821 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     5881 gggactttcc acaccctaac tgacacacat tccacagaat taattcgcgt taaatttttg
     5941 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6001 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6061 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6121 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6181 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6241 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
//
