LOCUS       sequence-382311-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-Creb5 fusion construct (N-terminus of Creb5,128aa, with T59/T61
            A mutation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  AUTHORS   Zhang CH, Gao Y, Jadhav U, Hung HH, Holton KM, Grodzinsky AJ, 
            Shivdasani RA, Lassar AB
  TITLE     Creb5 establishes the competence for Prg4 expression in articular 
            cartilage.
  JOURNAL   Commun Biol. 2021 Mar 12;4(1):332. doi: 10.1038/s42003-021-01857-0.
  PUBMED    33712729
REFERENCE   2  (bases 1 to 6197)
  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"; journalName: "Commun 
            Biol."; date: "2021-03-12"; volume: "4(1):332. doi"; pages: " 
            10.1038/s42003-021-01857-0"
COMMENT     Sequence Label: GAL4-Creb5 (1- 128) T59/T61 A
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..48)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     83..102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(162..178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(162..178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(175..197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    186..202
                     /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(210..216,217..234,235..240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    255..276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(393..410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(564..1152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(644..663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1323..2114,2115..2183))
                     /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     1946..1965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2184..2288)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2363..2382)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2554..2933
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2934..3137
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3088..3108
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3182..3201
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3182..3200
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3217..3657
                     /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(4069..4087)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4070..4087)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4107..4124)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4113..4337
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4383..4811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4470..4489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4680..4701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4820..4840)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4825..5154
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5005..5140
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5067..5086
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5221..6015
                     /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(5275..5294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5885..5904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6189..6197,1..113)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
       61 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
      121 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
      181 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
      241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      361 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1621 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     1681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     1741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     1801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     1861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     1921 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     1981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2281 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg
     2341 atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
     2401 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
     2461 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
     2521 tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat
     2581 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2641 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2701 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2761 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2821 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2881 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     2941 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     3001 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     3061 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     3121 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     3181 ttaatacgac tcactatagg gagacccaag cttaccatga agctactgtc ttctatcgaa
     3241 caagcatgcg atatttgccg acttaaaaag ctcaagtgct ccaaagaaaa accgaagtgc
     3301 gccaagtgtc tgaagaacaa ctgggagtgt cgctactctc ccaaaaccaa aaggtctccg
     3361 ctgactaggg cacatctgac agaagtggaa tcaaggctag aaagactgga acagctattt
     3421 ctactgattt ttcctcgaga agaccttgac atgattttga aaatggattc tttacaggat
     3481 ataaaagcat tgttaacagg attatttgta caagataatg tgaataaaga tgccgtcaca
     3541 gatagattgg cttcagtgga gactgatatg cctctaacat tgagacagca tagaataagt
     3601 gcgacatcat catcggaaga gagtagtaac aaaggtcaaa gacagttgac tgtatcgccg
     3661 gaattcatga tttatgagga atccaagatg aatttggagc aggagaggcc gtttgtctgc
     3721 agtgccccag gctgctccca gcgcttccca acagaggatc atctgatgat tcataggcac
     3781 aagcatgaaa tgactttgaa gtttccctca ataaaaacag acaatatgtt atcagatcaa
     3841 gctccagccc caacgagatt cctgaagaac tgcgaggagg tggggctctt cagtgagttg
     3901 gactgctccc tagagcatga gttcaggaag gctcaggagg aagagagcag caagcggaat
     3961 atctcgatgc ataaccccgt tggtggggcc atggcggggc ctggagctca ccagctcggc
     4021 agcgcccgga tgcccaacca tgacaccagc tgatctagag ggccctattc tatagtgtca
     4081 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4141 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4201 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4261 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4321 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4381 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4441 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4561 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4681 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4741 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4801 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     4861 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     4921 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     4981 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     5041 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     5101 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     5161 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     5221 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5281 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5341 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5401 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5461 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5521 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5581 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5641 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5701 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     5761 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5821 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5881 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5941 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6001 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6061 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6121 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6181 cccaccccaa cttgttt
//
