LOCUS       sequence-426589-        6280 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of Gal4-DBD OCT4 AroPERFECT N.
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-OCT4_AroPERFECT_N
FEATURES             Location/Qualifiers
     source          1..6280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             710..1150
                     /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(1599..1617)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1600..1617)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1637..1654)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1643..1867
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1913..2341
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2000..2019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2210..2231
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2350..2370)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2355..2684
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2535..2670
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2597..2616
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2751..3545
                     /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(2805..2824)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3415..3434
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3719..3840
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3756..3775)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3810..3829
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3889..3905)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3889..3905)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3902..3924)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3913..3929
                     /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(3937..3943,3944..3961,3962..3967))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3982..4003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4120..4137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4291..4879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4371..4390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5050..5841,5842..5910))
                     /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     5673..5692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5911..6015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6090..6109)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattca tgaagctact
      721 gtcttctatc gaacaagcat gcgatatttg ccgacttaaa aagctcaagt gctccaaaga
      781 aaaaccgaag tgcgccaagt gtctgaagaa caactgggag tgtcgctact ctcccaaaac
      841 caaaaggtct ccgctgacta gggcacatct gacagaagtg gaatcaaggc tagaaagact
      901 ggaacagcta tttctactga tttttcctcg agaagacctt gacatgattt tgaaaatgga
      961 ttctttacag gatataaaag cattgttaac aggattattt gtacaagata atgtgaataa
     1021 agatgccgtc acagatagat tggcttcagt ggagactgat atgcctctaa cattgagaca
     1081 gcatagaata agtgcgacat catcatcgga agagagtagt aacaaaggtc aaagacagtt
     1141 gactgtatcg gcgatcatgg ctggacacct ggcttcagac ttcgcctcac ccccaccagg
     1201 tgggggtgat gggtcagcag ggctgttcga gccgggcgtg gatcctcgaa ccctaagcca
     1261 agggcctcca tggggtgggc ctggaatcgg accaggctca gaggtattgg ggatctggtc
     1321 cccatgtccg cccgcagagt gcggagggat ggcatgtgga ttccctcagg ttggactggg
     1381 cctagtcccc caagttggcg tggagtacac tttgcagcct gagggccagg caggagcacg
     1441 agtggaaagc ttcaactcag agggaacctc ctctgagccc tgtgccgacc gcccctacaa
     1501 tgccgtgaag ttggagaagg tggaaccaac tcccgaggag ttttcccagg acatgaaagc
     1561 cctgcagaag gagctagaac aggtctagag ggccctattc tatagtgtca cctaaatgct
     1621 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1681 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1741 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1801 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1861 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     1921 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1981 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2041 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2101 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2161 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2221 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2281 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2341 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2401 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2461 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2521 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2581 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     2641 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     2701 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     2761 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     2821 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     2881 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     2941 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3001 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3061 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3121 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3181 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3241 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3301 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3361 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3421 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3481 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3541 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3601 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3661 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     3721 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3781 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3841 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     3901 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     3961 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4021 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4081 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4201 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4261 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4321 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4381 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4441 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4501 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4561 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4621 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4681 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4741 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4801 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4861 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4921 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4981 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5041 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5101 ttcatccata gttgcctgac ttcccgtcgt gtagataact acgatacggg agggcttacc
     5161 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5221 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5281 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5341 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5401 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5461 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5521 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5581 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5641 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5701 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5761 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5821 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5881 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5941 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6001 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6061 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6121 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6181 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     6241 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
