LOCUS       sequence-383808-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-Creb5 fusion construct (full length, 508aa).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  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 7337)
  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- 508)
FEATURES             Location/Qualifiers
     source          1..7337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1800..1819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1991..2370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2371..2574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2525..2545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2619..2638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2619..2637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2654..3094
                     /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(4646..4664)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4647..4664)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4684..4701)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4690..4914
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4960..5388
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5047..5066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5257..5278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5397..5417)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5402..5731
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5582..5717
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5644..5663
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5798..6592
                     /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(5852..5871)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6462..6481
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6766..6887
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6803..6822)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6857..6876
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6936..6952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6936..6952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6949..6971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6960..6976
                     /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(6984..6990,6991..7008,7009..7014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7029..7050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7167..7184)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     1801 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1861 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1921 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1981 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2581 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2641 acccaagctt accatgaagc tactgtcttc tatcgaacaa gcatgcgata tttgccgact
     2701 taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg
     2761 ggagtgtcgc tactctccca aaaccaaaag gtctccgctg actagggcac atctgacaga
     2821 agtggaatca aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagaaga
     2881 ccttgacatg attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt
     2941 atttgtacaa gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac
     3001 tgatatgcct ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag
     3061 tagtaacaaa ggtcaaagac agttgactgt atcgccggaa ttcatgattt atgaggaatc
     3121 caagatgaat ttggagcagg agaggccgtt tgtctgcagt gccccaggct gctcccagcg
     3181 cttcccaaca gaggatcatc tgatgattca taggcacaag catgaaatga ctttgaagtt
     3241 tccctcaata aaaacagaca atatgttatc agatcaaact ccaaccccaa cgagattcct
     3301 gaagaactgc gaggaggtgg ggctcttcag tgagttggac tgctccctag agcatgagtt
     3361 caggaaggct caggaggaag agagcagcaa gcggaatatc tcgatgcata accccgttgg
     3421 tggggccatg gcggggcctg gagctcacca gctcggcagc gcccggatgc ccaaccatga
     3481 caccagcgtt gtgattcagc aagccatgcc gtcaccccag tccagctctg tcatcacgca
     3541 ggccccttcc accaaccgcc agatcgggcc tgtcccaggc tctctatctt ctctgctcca
     3601 tcttcacaac agacagagac agcccatgcc agcctccatg cctgggaccc tgcccaaccc
     3661 tacaatgccg ggctcttctg ccgtcttgat gccaatggag agacaaatgt cagtgaactc
     3721 caacctcctg ggaatgcaag gtccaaatct cagcaacccc tgtgcttctc cccaagtcca
     3781 gccaatgcat tcagaagcta agatgaggct gaaggctgca ttgactcacc accctgctgc
     3841 catgtcgaat gggaatatga acaccatggg ccacatgatg gaaatgatgg gctcccggca
     3901 ggaccagacg ccacaccatc acatgcactc acacccgcat cagcaccaga cactgccggc
     3961 ccaccacccc tacccgcacc agcaccagca cccagcacac caccctcatc ctcaacccca
     4021 tcaccagcag aaccacccac atcatcactc ccactcccac cttcatgcac acccagcaca
     4081 tcaccagacc tcgccacacc cacccctgca ctccggcacc caagcacagg tttcaccagc
     4141 aacacaacag atgcagccca cccagacaat acagccgccc cagcccacag gggggcgccg
     4201 gcgaagggtg gtggatgagg atcctgacga gaggcggcgg aaatttctgg aacggaaccg
     4261 ggcggccgcc acccgctgca ggcagaagag gaaggtctgg gtgatgtcac tggaaaagaa
     4321 agcagaagag ctcacccaga caaacatgca gcttcagaac gaagtgtcca tgttgaagaa
     4381 cgaggtggca cagctgaagc agctgttgtt aacacataaa gactgcccca taacagccat
     4441 gcagaaagaa tcacaaggat atctcagtcc agagagtagc cctcctgcaa gtcccacccc
     4501 cgcatgctca cagcagcagg tcatccagca taacaccatc accacttcct cggcagtcag
     4561 cgaggtggta ggaagctcta ccctcagcca gctcaccact cacagaacag acctgaaccc
     4621 catcctctga tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     4681 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4741 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4801 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4861 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4921 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4981 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5041 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5101 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5161 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5221 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5281 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5341 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     5401 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5461 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5521 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5581 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5641 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5701 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5761 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5821 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5881 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5941 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     6001 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     6061 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     6121 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     6181 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6241 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6301 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     6361 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6421 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6481 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6541 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6601 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6661 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6721 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6781 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6841 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6901 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6961 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     7021 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     7081 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     7141 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     7201 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7261 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7321 gccgcgttgc tggcgtt
//
