LOCUS       sequence-430414-        6279 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD HOXD4 AroLITE S.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6279)
  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 6279)
  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-AroLITE_S
FEATURES             Location/Qualifiers
     source          1..6279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      476..1064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     965..984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1218..1235
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1352..1373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1388..1393,1394..1411,1412..1418)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1426..1442
                     /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     1431..1453
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1450..1466
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1450..1466
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1515..1636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1526..1545)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1580..1599
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1810..2604)
                     /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(1921..1940)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2531..2550
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2671..3000)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2685..2820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2739..2758)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2985..3005
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(3014..3442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3336..3355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3488..3712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3701..3718
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3738..3756
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3738..3755
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4204..4644)
                     /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(4706..4725)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4707..4725)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4770..4973)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4799..4819)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4974..5353
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5525..5544
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5619..5723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5724..5792,5793..6279,1..305)
                     /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(5942..5961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
       61 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
      121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
      181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg ggaagtcagg
      241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
      301 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
      361 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
      421 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      481 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      541 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      601 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      661 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      721 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      781 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      841 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      901 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      961 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1021 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1081 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1141 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     1201 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     1261 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1321 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     1381 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1441 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
     1501 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1561 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1621 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     1681 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     1741 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     1801 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     1861 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     1921 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     1981 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     2041 atgggtcacg acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc
     2101 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     2161 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     2221 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     2281 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     2341 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     2401 cgatagccgc gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac
     2461 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     2521 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     2581 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatcc
     2641 gaaaatggat atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc
     2701 tcctcactac ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa
     2761 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
     2821 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
     2881 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
     2941 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
     3001 attccacaga attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3061 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3121 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3181 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3241 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3301 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3361 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3421 aatgcgccgc tacagggcgc gtggggatac cccctagagc cccagctggt tctttccgcc
     3481 tcagaagcca tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc
     3541 cccttgctgt cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc
     3601 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
     3661 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
     3721 gatcagcgag ctctagcatt taggtgacac tatagaatag ggccctctag acacaacttt
     3781 cttcatagac ggagacacaa ccgcaggttg tttaagggca gttccactcg gtggttgttt
     3841 aggatcgctt tggctggacg ctcgtgcccc gggcaacggg gctggtgggg gagcgggcgg
     3901 tgcaggacat ggttctcctg gtgctgcact atgtccacct ggcccgccgg gttcttggcc
     3961 atgtccgcgg gctgggagtg ctgatccagg tcccggcccg gacccgccgg atggttgttc
     4021 gcctgaatct ggcctgggag aaaggcctgg tggttgggaa tctgcccctt gtgcaccacc
     4081 accataagaa tcagcacctt gttctccgag ggatccgcct tgcagggatt cttcacaagg
     4141 cggggatttt ggatctacgg atttgctatt gaccatactg ctgctcatca ccatatgtat
     4201 cgccgataca gtcaactgtc tttgaccttt gttactactc tcttccgatg atgatgtcgc
     4261 acttattcta tgctgtctca atgttagagg catatcagtc tccactgaag ccaatctatc
     4321 tgtgacggca tctttattca cattatcttg tacaaataat cctgttaaca atgcttttat
     4381 atcctgtaaa gaatccattt tcaaaatcat gtcaaggtct tctcgaggaa aaatcagtag
     4441 aaatagctgt tccagtcttt ctagccttga ttccacttct gtcagatgtg ccctagtcag
     4501 cggagacctt ttggttttgg gagagtagcg acactcccag ttgttcttca gacacttggc
     4561 gcacttcggt ttttctttgg agcacttgag ctttttaagt cggcaaatat cgcatgcttg
     4621 ttcgatagaa gacagtagct tcatgaattc cagcacactg gcggccgtta ctagtggatc
     4681 cgagctcggt accaagcttg ggtctcccta tagtgagtcg tattaatttc gataagccag
     4741 taagcagtgg gttctctagt tagccagaga gctctgctta tatagacctc ccaccgtaca
     4801 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     4861 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     4921 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     4981 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     5041 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     5101 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     5161 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     5221 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     5281 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     5341 aataatcaat gtcaacgcgt atatctggcc cgtacatcgc gaagcagcgc aaaacgccta
     5401 accctaagca gattcttcat gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt
     5461 ttgctcgcgc actactcagc gacctccaac acacaagcag ggagcagata ctggcttaac
     5521 tatgcggcat cagagcagat tgtactgaga gtgcaccata ggggatcggg agatctcccg
     5581 atccgtcgac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5641 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5701 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5761 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5821 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5881 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5941 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     6001 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6061 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6121 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6181 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6241 acgacgagcg tgacaccacg atgcctgtag caatggcaa
//
