LOCUS       sequence-382313-        6197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GAL4-Creb5 fusion construct (N-terminus of Creb5,128aa, with C18/C23
            S 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) C18/C23 S
FEATURES             Location/Qualifiers
     source          1..6197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(132..720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(212..231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(891..1682,1683..1751))
                     /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     1514..1533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1752..1856)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1931..1950)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2122..2501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2502..2705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2656..2676
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2750..2769
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2750..2768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2785..3225
                     /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(3637..3655)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3638..3655)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3675..3692)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3681..3905
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3951..4379
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4038..4057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4248..4269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4388..4408)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4393..4722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4573..4708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4635..4654
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4789..5583
                     /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(4843..4862)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5453..5472
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5757..5878
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5794..5813)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5848..5867
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5927..5943)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5927..5943)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5940..5962)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5951..5967
                     /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(5975..5981,5982..5999,6000..6005))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6020..6041
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6158..6175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
       61 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      301 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      541 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      661 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      721 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      781 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1021 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1861 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1921 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1981 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     2041 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     2101 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     2161 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2221 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2281 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2341 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2401 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2461 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2521 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2581 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     2641 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2701 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2761 actataggga gacccaagct taccatgaag ctactgtctt ctatcgaaca agcatgcgat
     2821 atttgccgac ttaaaaagct caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg
     2881 aagaacaact gggagtgtcg ctactctccc aaaaccaaaa ggtctccgct gactagggca
     2941 catctgacag aagtggaatc aaggctagaa agactggaac agctatttct actgattttt
     3001 cctcgagaag accttgacat gattttgaaa atggattctt tacaggatat aaaagcattg
     3061 ttaacaggat tatttgtaca agataatgtg aataaagatg ccgtcacaga tagattggct
     3121 tcagtggaga ctgatatgcc tctaacattg agacagcata gaataagtgc gacatcatca
     3181 tcggaagaga gtagtaacaa aggtcaaaga cagttgactg tatcgccgga attcatgatt
     3241 tatgaggaat ccaagatgaa tttggagcag gagaggccgt ttgtcagcag tgccccaggc
     3301 agctcccagc gcttcccaac agaggatcat ctgatgattc ataggcacaa gcatgaaatg
     3361 actttgaagt ttccctcaat aaaaacagac aatatgttat cagatcaaac tccaacccca
     3421 acgagattcc tgaagaactg cgaggaggtg gggctcttca gtgagttgga ctgctcccta
     3481 gagcatgagt tcaggaaggc tcaggaggaa gagagcagca agcggaatat ctcgatgcat
     3541 aaccccgttg gtggggccat ggcggggcct ggagctcacc agctcggcag cgcccggatg
     3601 cccaaccatg acaccagctg atctagaggg ccctattcta tagtgtcacc taaatgctag
     3661 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3721 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3781 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3841 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3901 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3961 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4021 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4081 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4141 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4201 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4261 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4321 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4381 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4441 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4501 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4561 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4621 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4681 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4741 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4801 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4861 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4921 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4981 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5041 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5101 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5161 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5221 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5281 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5341 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5401 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5461 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5521 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5581 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5641 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5701 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5761 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5821 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5881 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5941 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6001 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6061 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6121 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6181 cggtcgttcg gctgcgg
//
