LOCUS       sequence-310831-        5052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of QFGal4 mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5052)
  AUTHORS   Burgess J, Burrows JT, Sadhak R, Chiang S, Weiss A, D'Amata C, 
            Molinaro AM, Zhu S, Long M, Hu C, Krause HM, Pearson BJ
  TITLE     An optimized QF-binary expression system for use in zebrafish.
  JOURNAL   Dev Biol. 2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007.
  PUBMED    32697972
REFERENCE   2  (bases 1 to 5052)
  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: "Dev Biol. 
            2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007."
COMMENT     Sequence Label: pCS2-QFGal4-SV40pA
FEATURES             Location/Qualifiers
     source          1..5052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        120..1104
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1139..1157
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1139..1156
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1833..2177
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     polyA_signal    2208..2342
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2232..2251)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2286..2305
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2455..2475)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2455..2473)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2494..2510)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2494..2510)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2507..2529)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2518..2534
                     /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(2542..2548,2549..2566,2567..2572))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2587..2608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2725..2742)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2896..3484)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2976..2995)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3655..4446,4447..4515))
                     /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     4278..4297
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4516..4620)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(4646..5052,1..49))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4783..4804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4995..5014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
       61 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
      121 tcgaccgcca attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg
      181 acctgtgcca attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg
      241 accccagcca attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg
      301 cactgtgcca actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg
      361 gccctgtgcc aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg
      421 ctatatgcca ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat
      481 tggccaggtt caatactatg tattggccct atgccatata gtattccata tatgggtttt
      541 cctattgacg tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc
      601 taataccgcc catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct
      661 attgacgtca tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt
      721 acggtaaatg gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga
      781 cgtcaatggg atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa
      841 tgacggtaaa tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta
      901 cattactatt ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg
      961 taaatggccc gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg
     1021 acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat
     1081 ataagcaatg ctcgtttagg gaaccgccat tctgcctggg gacgtcggag caagcttgat
     1141 ttaggtgaca ctatagaata caagctactt gttctttttg caggatcctc gccgccacca
     1201 tgccgcctaa acgcaagaca ctcaatgccg ctgccgaagc caatgcccac gctgatggcc
     1261 atgctgatgg caatgctgat ggtcatgtcg ctaacactgc agcaagcagc aacaacgccc
     1321 gttttgcgga cttgaccaac attgacacac ccggcctcgg ccctaccacg acgacgttac
     1381 ttgtcgagcc cgctcgttcg aaacgccaga gagtctcgag ggcctgtgat cagtgtcgag
     1441 ctgcacgtga aaagtgtgat ggaatccagc cggcttgctt cccctgtgtg tcgcagggcc
     1501 ggtcgtgtac ctaccaggcc agtcccaaga agcgaggagt ccagacgggc tacatccgca
     1561 ctctcgaact ggctctggct tggatgttcg agaacgttgc ccgcagcgag gacgccctcc
     1621 acaatctttt ggtccgtgat gctggccagg gcagcgctct cctggtcggc aaagactcgc
     1681 ctgctgcaga acgcctgcat gcaagatggg cgacgagtcg agtcaacaaa agcatcaccc
     1741 gtcttctctc aggtgctagc actagtagcg ccggtcaaca gcaacagctg caatcattag
     1801 tgccactgac cccgtctgct ttgtttggtg gcgccaattt taatcaaagt gggaatattg
     1861 ctgatagctc attgtccttc actttcacta acagtagcaa cggtccgaac ctcataacaa
     1921 ctcaaacaaa ttctcaagcg ctttcacaac caattgcctc ctctaacgtt catgataact
     1981 tcatgaataa tgaaatcacg gctagtaaaa ttgatgatgg taataattca aaaccactgt
     2041 cacctggttg gacggaccaa actgcgtata acgcgtttgg aatcactaca gggatgttta
     2101 ataccactac aatggatgat gtatataact atctattcga tgatgaagat accccaccaa
     2161 acccaaaaaa agagtaaatc tagaactata gtgagtcgta ttacgtagat ccagacatga
     2221 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2281 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2341 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2401 tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta
     2461 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     2521 ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta aagcctgggg
     2581 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     2641 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     2701 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2761 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2821 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2881 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2941 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3001 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3061 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     3121 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3181 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3241 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3301 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3361 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3421 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3481 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3541 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3601 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3661 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3721 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3781 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3841 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3901 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3961 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     4021 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     4081 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     4141 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4201 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4261 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4321 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4381 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4441 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4501 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4561 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     4621 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
     4681 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     4741 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     4801 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     4861 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     4921 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     4981 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     5041 gcggtcacgc tg
//
