LOCUS       sequence-382307-        6196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The control vector GAL4 (1-147) was generated by introducing a stop 
            codon immediately after the GAL4 (1-147) sequence in the GAL4-Creb5 
            (1-128) vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6196)
  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 6196)
  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(1-147)
FEATURES             Location/Qualifiers
     source          1..6196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    183..304
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(220..239)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     274..293
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(353..369)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(353..369)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(366..388)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    377..393
                     /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(401..407,408..425,426..431))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    446..467
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(584..601)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(755..1343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(835..854)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1514..2305,2306..2374))
                     /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     2137..2156
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2375..2479)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2554..2573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2745..3124
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3125..3328
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3279..3299
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3373..3392
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3373..3391
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3408..3848
                     /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(4259..4277)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4260..4277)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4297..4314)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4303..4527
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4573..5001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4660..4679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4870..4891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5010..5030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5015..5344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5195..5330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5257..5276
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(5411..6196,1..9)
                     /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(5465..5484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6075..6094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
       61 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
      121 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
      181 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      241 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      301 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
      361 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
      421 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
      481 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
      541 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
      601 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      661 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      721 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      781 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      841 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      901 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      961 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1021 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1081 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1141 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1201 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1261 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1321 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1381 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1441 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1501 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1561 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1621 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1681 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1741 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1801 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1861 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1921 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1981 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2041 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2101 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2161 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2221 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2281 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2341 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2401 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2461 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     2521 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     2581 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     2641 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     2701 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
     2761 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     2821 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     2881 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     2941 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     3001 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     3061 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     3121 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     3181 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     3241 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     3301 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
     3361 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg aagctactgt
     3421 cttctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc tccaaagaaa
     3481 aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct cccaaaacca
     3541 aaaggtctcc gctgactagg gcacatctga cagaagtgga atcaaggcta gaaagactgg
     3601 aacagctatt tctactgatt tttcctcgag aagaccttga catgattttg aaaatggatt
     3661 ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat gtgaataaag
     3721 atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca ttgagacagc
     3781 atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa agacagttga
     3841 ctgtatcgcc ggaattctga tttatgagga atccaagatg aatttggagc aggagaggcc
     3901 gtttgtctgc agtgccccag gctgctccca gcgcttccca acagaggatc atctgatgat
     3961 tcataggcac aagcatgaaa tgactttgaa gtttccctca ataaaaacag acaatatgtt
     4021 atcagatcaa actccaaccc caacgagatt cctgaagaac tgcgaggagg tggggctctt
     4081 cagtgagttg gactgctccc tagagcatga gttcaggaag gctcaggagg aagagagcag
     4141 caagcggaat atctcgatgc ataaccccgt tggtggggcc atggcggggc ctggagctca
     4201 ccagctcggc agcgcccgga tgcccaacca tgacaccagc tgatctagag ggccctattc
     4261 tatagtgtca cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc
     4321 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     4381 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     4441 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4501 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4561 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4621 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4681 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4741 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4801 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4861 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4921 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4981 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5041 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5101 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5161 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5221 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5281 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5341 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     5401 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5461 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5521 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5581 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5641 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5701 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5761 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5821 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5881 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5941 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6001 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6061 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6121 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6181 tcgccttctt gacgag
//
