LOCUS       sequence-334444-        4381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for synthesis of GFP chimeric RNA that contained vasa-3'UTRs
            of zebrafish by in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4381)
  AUTHORS   Yoshizaki G, Tago Y, Takeuchi Y, Sawatari E, Kobayashi T, Takeuchi T
  TITLE     Green fluorescent protein labeling of primordial germ cells using a 
            nontransgenic method and its application for germ cell 
            transplantation in salmonidae.
  JOURNAL   Biol Reprod. 2005 Jul;73(1):88-93. doi: 
            10.1095/biolreprod.104.034249. Epub 2005 Mar 2.
  PUBMED    15744027
REFERENCE   2  (bases 1 to 4381)
  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"; doi: 
            "10.1095/biolreprod.104.034249"; journalName: "Biol Reprod"; date: 
            "2005-07"; volume: "73"; issue: "1"; pages: "88-93"
COMMENT     Sequence Label: Gfp-zf-vasa
FEATURES             Location/Qualifiers
     source          1..4381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             46..765
                     /codon_start=1
                     /product="humanized Renilla green fluorescent protein"
                     /label=hrGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKQILKNTGLQEIMSFKVNLEGVVNNHVFTMEGCGKGNILFGN
                     QLVQIRVTKGAPLPFAFDILSPAFQYGNRTFTKYPEDISDFFIQSFPAGFVYERTLRYE
                     DGGLVEIRSDINLIEEMFVYRVEYKGRNFPNDGPVMKKTITGLQPSFEVVYMNDGVLVG
                     QVILVYRLNSGKFYSCHMRTLMKSKGVVKDFPEYHFIQHRLEKTYVEDGGFVEQHETAI
                     AQLTSLGKPLGSLHEWV"
     promoter        complement(1523..1541)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1524..1541)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1559..1575)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1559..1575)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1572..1594)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1583..1599
                     /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(1607..1613,1614..1631,1632..1637))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1652..1673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1790..1807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1961..2549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2041..2060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2720..3511,3512..3580))
                     /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     3343..3362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3581..3685)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3763..4218)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3900..3921)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4112..4131
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4344..4366
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4358..4375
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4359..4375
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggc ccgacgtcca caaccatggt gagcaagcag
       61 atcctgaaga acaccggcct gcaggagatc atgagcttca aggtgaacct ggagggcgtg
      121 gtgaacaacc acgtgttcac catggagggc tgcggcaagg gcaacatcct gttcggcaac
      181 cagctggtgc agatccgcgt gaccaagggc gcccccctgc ccttcgcctt cgacatcctg
      241 agccccgcct tccagtacgg caaccgcacc ttcaccaagt accccgagga catcagcgac
      301 ttcttcatcc agagcttccc cgccggcttc gtgtacgagc gcaccctgcg ctacgaggac
      361 ggcggcctgg tggagatccg cagcgacatc aacctgatcg aggagatgtt cgtgtaccgc
      421 gtggagtaca agggccgcaa cttccccaac gacggccccg tgatgaagaa gaccatcacc
      481 ggcctgcagc ccagcttcga ggtggtgtac atgaacgacg gcgtgctggt gggccaggtg
      541 atcctggtgt accgcctgaa cagcggcaag ttctacagct gccacatgcg caccctgatg
      601 aagagcaagg gcgtggtgaa ggacttcccc gagtaccact tcatccagca ccgcctggag
      661 aagacctacg tggaggacgg cggcttcgtg gagcagcacg agaccgccat cgcccagctg
      721 accagcctgg gcaagcccct gggcagcctg cacgagtggg tgtaatagcc gcgggaattc
      781 gattgtcgac tgggaataac tggcctcaca cctgttatat ttattttatt tttattttat
      841 ttgtgcgagc ttcttcaata ccatttcagg atatttgtta atgttgaatg actaaagtag
      901 cattaggata aagctgttgt atttagagtc acattgaatg ctgtaggttt gacctcaagg
      961 acgataatca agactaatac cttttggcag tttgatttga tcatcttttt ggtatcactc
     1021 gtttggactg gagcacaact gaattagtgg atttgtttta ctagagcaca agtgcagaag
     1081 tgggtgggtt tgtttaatca cattttaaca ctttttgcta ggagtgtatt gagttttgta
     1141 agctttgacc taaaggtgtt tcctacagtt aaatctatta atcagttcta ttgttgattg
     1201 tgactgtttg taggttattt ttcctgacat ttccgtcttg aagccagagt atagaacaag
     1261 aaacttgttt tgcagatgta ccttgccaga taaaggtagc taaagtcatt gtttaacaga
     1321 catctcttta attttaaagt caaaatgttg ataaaattca gtcaagaggc agtgactgac
     1381 tgtaacagag cggtgcttcc tcactagctg aagtggctca aaataaagac ggatactggt
     1441 gaaatcacta gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc
     1501 gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt
     1561 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1621 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1681 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1741 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1801 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1861 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1921 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1981 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     2041 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     2101 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     2161 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     2221 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     2281 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     2341 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     2401 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2461 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2521 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2581 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2641 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2701 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2761 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2821 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2881 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2941 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     3001 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     3061 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     3121 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     3181 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     3241 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     3301 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     3361 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3421 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3481 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3541 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3601 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3661 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt
     3721 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata
     3781 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3841 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3901 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3961 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4021 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4081 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4141 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4201 cgccgctaca gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     4261 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     4321 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
     4381 g
//
