LOCUS       sequence-234981-        5995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent vector with zebrafish pax3a enhancer 5 (strong 
            activity).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5995)
  AUTHORS   Chong-Morrison V, Mayes S, Simoes FC, Senanayake U, Carroll DS, 
            Riley PR, Wilson SW, Sauka-Spengler T
  TITLE     Ac/Ds transposition for CRISPR/dCas9-SID4x epigenome modulation in 
            zebrafish.
  JOURNAL   Biol Open. 2023 Jun 15;12(6):bio059995. doi: 10.1242/bio.059995. 
            Epub 2023 Jun 27.
  PUBMED    37367831
REFERENCE   2  (bases 1 to 5995)
  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: "Biol 
            Open."; date: "2023-06-15"; volume: "12(6):bio059995. doi"; pages: "
            10.1242/bio.059995. Epub 2023 Jun 27"
COMMENT     Sequence Label: pVC-Ds-Pax3a_enh5-E1b:eGFP-Ds
FEATURES             Location/Qualifiers
     source          1..5995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..86
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     78..95
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     79..95
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     105..124
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        105..123
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     intron          1189..1761
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     regulatory      1835..1844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1841..1843,1844..1846,1847..2560)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1841..1843,1844..1846,1847..2557)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1886..1907)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2147..2166)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2494..2515
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2770..2851
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2807..2826)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2861..2880
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3286..3306)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3286..3304)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3325..3341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3325..3341)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3338..3360)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3349..3365
                     /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(3373..3379,3380..3397,3398..3403))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3418..3439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3556..3573)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3727..4315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3807..3826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4486..5277,5278..5346))
                     /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     5109..5128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5347..5451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5477..5932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5614..5635)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5826..5845
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
       61 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat
      121 agggcgaatt gggtacccga cgcagggatg aaagtaggat gggaaaatcc cgtaccgacc
      181 gttatcgtat aaccgatttt gttagtttta tcccgatcga tttcgaaccc gaggtaaaaa
      241 acgaaaacgg aacggaaacg ggatatacaa aacggtaaac ggaaacggaa acggtagagc
      301 tagtttcccg accgtttcac cgggatcccg tttttaatcg ggatgatccc gtttcgttac
      361 cgtattttct aattcgggat gactgcaaca gatccctcga ctcgagttta cgtaccgcta
      421 gccacacaat tgtcccaaca caaatcgatt aaattaaaat aatcgtttta tttttaatcc
      481 acttcaattt gtaaaaacaa ttaattaaaa aagtagtttg aacaagcagc aaaagtaata
      541 ttttcagtat acaaagcgcc aaaagcactg cagataattt cttatttgtt tttgcgccct
      601 cttgcggaca gaagaatatt gcagggaaag ccgctttgag aggtgcgtga atcttcatgt
      661 ctctccggct ttaccacctg cagagaggag cgctactggt agccatgatc acctgagagc
      721 aataatgccg gagctcaggc cccggggcca aggttcaccg cgggggcaat aaactcctgt
      781 ccccctgcca tcatgtaaag acattaccgc gccctgcggc cctcacaaca aacactccat
      841 ttccaaaggc tctctgccgc ttactgggtc gctttattta ccctgtgtca tgcaaaagtc
      901 ttaatttcct ttgaatccca gtgctgctgt attgccttga gttacttaca ttcattacca
      961 ccgtttaatt agtaccatca aacgcttcac ggtttacagt ggctcaatgt gacattagtc
     1021 gagactgtta aactcattag caaatactga taagggggta aaggtgagat ggctgctagc
     1081 aagcttgcgg cgatatcgac tctagagggt atataatgga tcccatcgcg tctcagcctc
     1141 actttgagct cctccacacg aatttcgacc gatcctgaga acttcagggt gagtttgggg
     1201 acccttgatt gttctttctt tttcgctatt gtaaaattca tgttatatgg agggggcaaa
     1261 gttttcaggg tgttgtttag aatgggaaga tgtcccttgt atcaccatgg accctcatga
     1321 taattttgtt tctttcactt tctactctgt tgacaaccat tgtctcctct tattttcttt
     1381 tcattttctg taactttttc gttaaacttt agcttgcatt tgtaacgaat ttttaaattc
     1441 acttttgttt atttgtcaga ttgtaagtac tttctctaat cacttttttt tcaaggcaat
     1501 cagggtatat tatattgtac ttcagcacag ttttagagaa caattgttat aattaaatga
     1561 taaggtagaa tatttctgca tataaattct ggctggcgtg gaaatattct tattggtaga
     1621 aacaactaca ccctggtcat catcctgcct ttctctttat ggttacaatg atatacactg
     1681 tttgagatga ggataaaata ctctgagtcc aaaccgggcc cctctgctaa ccatgttcat
     1741 gccttcttct ctttcctaca gctcctgggc aacgtgctgg ttgttgtgct gtctcatcat
     1801 tttggcaaag aattcctcga cggatccacc ggtcgccacc atggtgagca agggcgagga
     1861 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1921 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1981 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2041 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2101 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2161 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2221 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2281 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2341 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2401 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2461 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2521 cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg actctagaat
     2581 tccaactgag cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggcct
     2641 actagtaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2701 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2761 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2881 aatgtagagt agacttatat ggcttcttat gttagccaag agcccaagac ttatcactta
     2941 tgtgctacat taaactatgt gtgctccaga tttatatgga ttttatctat gtttaattaa
     3001 gacttgtgtt tacaattttt tatatttgtt tttaagtttt gaatatatgt tttcatgtgt
     3061 gattttaccg aacaaaaata ccggttcccg tccgatttcg actttaaccc gaccggatcg
     3121 tatcggtttt cgattaccgt atttatcccg ttcgttttcg ttaccggtat atcccgtttt
     3181 cgtttccgtc ccgcaagtta aatatgaaaa tgaaaacggt agaggtattt taccgaccgt
     3241 taccgaccgt tttcatccct agtcggggag ctccagcttt tgttcccttt agtgagggtt
     3301 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3361 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3421 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3481 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3541 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3601 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3661 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3721 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3781 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3841 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3901 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3961 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4021 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4081 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4141 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4201 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4261 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4321 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4381 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4441 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4501 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4561 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4621 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4681 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4741 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4801 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4861 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4921 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4981 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5041 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5101 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5161 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5221 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5281 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5341 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5401 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5461 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5521 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5581 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5641 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5701 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5761 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5821 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5881 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     5941 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctat
//
