LOCUS       sequence-392212-        5803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of stdPCP-PH in zebrafish embryos.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5803)
  AUTHORS   Liang Y, Xu H, Cheng T, Fu Y, Huang H, Qian W, Wang J, Zhou Y, Qian 
            P, Yin Y, Xu P, Zou W, Chen B
  TITLE     Gene activation guided by nascent RNA-bound transcription factors.
  JOURNAL   Nat Commun. 2022 Nov 28;13(1):7329. doi: 10.1038/s41467-022-35041-7.
  PUBMED    36443367
REFERENCE   2  (bases 1 to 5803)
  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: "Nat 
            Commun."; date: "2022-11-28"; volume: "13(1):7329. doi"; pages: " 
            10.1038/s41467-022-35041-7"
COMMENT     Sequence Label: pCA31-pCS2-CMV-stdPCP-p65-HSF1
FEATURES             Location/Qualifiers
     source          1..5803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..17)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1..17)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(14..36)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    25..41
                     /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(49..55,56..73,74..79))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    94..115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(232..249)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(403..991)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(483..502)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1162..1953,1954..2022))
                     /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     1785..1804
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2023..2127)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2153..2608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2290..2311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2502..2521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2679..3663
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        3698..3716
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3698..3715
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4547..5089
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             5114..5485
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     polyA_signal    5518..5652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5542..5561)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5596..5615
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5765..5785)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5765..5783)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
       61 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
      121 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
      181 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
      241 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
      301 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
      361 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      421 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      481 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      541 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      601 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      661 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      721 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      781 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      841 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      901 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      961 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1021 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1081 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1141 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1201 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1261 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1321 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1381 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1441 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1501 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1561 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1621 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1681 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1741 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1801 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1861 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1921 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1981 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     2041 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     2101 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     2161 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     2221 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2281 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2341 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2401 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2461 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2521 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2581 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     2641 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
     2701 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     2761 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     2821 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
     2881 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     2941 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     3001 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     3061 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     3121 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     3181 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     3241 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     3301 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     3361 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     3421 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
     3481 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     3541 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     3601 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     3661 aaccgccatt ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac
     3721 aagctacttg ttctttttgc aggatcccat cgattcgaat tcaaggcctc tcgaggccac
     3781 catgttggct tccaaaacta tcgttctgag cgtaggtgaa gcgaccagaa cgttaactga
     3841 aattcaatct acagcagaca ggcaaatatt tgaagagaaa gtcggtccat tagtagggag
     3901 gttaagattg acagctagct taagacaaaa tggagctaaa actgcataca gggtcaattt
     3961 aaaattggat caagcagatg tcgttgatag cggtctacct aaggttagat acacgcaagt
     4021 gtggtcacac gatgtcacta ttgtcgcgaa ttctaccgaa gctagtagaa aatcattgta
     4081 tgatttaaca aaatccttgg tggctacctc tcaggtggaa gatcttgttg tgaatttagt
     4141 tccattaggt agggcggatc cgctagcctc caaaaccatc gttctttcgg tcggcgaggc
     4201 tactcgcact ctgactgaga tccagtccac cgcagaccgt cagatcttcg aagagaaggt
     4261 cgggcctctg gtgggtcggc tgcgcctcac ggcttcgctc cgtcaaaacg gagccaagac
     4321 cgcgtatcgc gtcaacctaa aactggatca ggcggacgtc gttgattccg gacttccgaa
     4381 agtgcgctac actcaggtat ggtcgcacga cgtgacaatc gttgcgaata gcaccgaggc
     4441 ctcgcgcaaa tcgttgtacg atttgaccaa gtccctcgtc gcgacctcgc aggtcgaaga
     4501 tcttgtcgtc aaccttgtgc cgctgggccg tccaccggtc gccacccctt cagggcagat
     4561 cagcaaccag gccctggctc tggcccctag ctccgctcca gtgctggccc agactatggt
     4621 gccctctagt gctatggtgc ctctggccca gccacctgct ccagcccctg tgctgacccc
     4681 aggaccaccc cagtcactga gcgctccagt gcccaagtct acacaggccg gcgaggggac
     4741 tctgagtgaa gctctgctgc acctgcagtt cgacgctgat gaggacctgg gagctctgct
     4801 ggggaacagc accgatcccg gagtgttcac agatctggcc tccgtggaca actctgagtt
     4861 tcagcagctg ctgaatcagg gcgtgtccat gtctcatagt acagccgaac caatgctgat
     4921 ggagtacccc gaagccatta cccggctggt gaccggcagc cagcggcccc ccgaccccgc
     4981 tccaactccc ctgggaacca gcggcctgcc taatgggctg tccggagatg aagacttctc
     5041 aagcatcgct gatatggact ttagtgccct gctgtcacag atttcctcta gtgggcaggg
     5101 aggaggtgga agcggcttca gcgtggacac cagtgccctg ctggacctgt tcagcccctc
     5161 ggtgaccgtg cccgacatga gcctgcctga ccttgacagc agcctggcca gtatccaaga
     5221 gctcctgtct ccccaggagc cccccaggcc tcccgaggca gagaacagca gcccggattc
     5281 agggaagcag ctggtgcact acacagcgca gccgctgttc ctgctggacc ccggctccgt
     5341 ggacaccggg agcaacgacc tgccggtgct gtttgagctg ggagagggct cctacttctc
     5401 cgaaggggac ggcttcgccg aggaccccac catctccctg ctgacaggct cggagcctcc
     5461 caaagccaag gaccccactg tctcctaatc tagaactata gtgagtcgta ttacgtagat
     5521 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5581 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5641 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     5701 tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     5761 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atg
//
