LOCUS       sequence-334445-        4346 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for synthesis of GFP chimeric RNA that contained vasa-3'UTRs
            of rainbow trout by in vitro transcription..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4346)
  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 4346)
  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-rt-vasa
FEATURES             Location/Qualifiers
     source          1..4346
                     /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(1488..1506)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1489..1506)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1524..1540)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1524..1540)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1537..1559)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1548..1564
                     /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(1572..1578,1579..1596,1597..1602))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1617..1638
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1755..1772)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1926..2514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2006..2025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2685..3476,3477..3545))
                     /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     3308..3327
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3546..3650)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3728..4183)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3865..3886)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4077..4096
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4309..4331
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4323..4340
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4324..4340
                     /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 gatttgggag tgatgatgca catctacata ttcttttagt ttttgttttc aagtcatgtt
      841 ttggttcgat ttaacttttt cttttagtgt tattttgttt aaagaaaata gttttgcact
      901 gcttgatatt tctggaaatg tgcgtgtggt cctggatatg ggaaaaccca ttaatgtcaa
      961 ccactggtta aaatctgtgt gcttgaggtg aaacttcaca aatgaatgtt ctagctatta
     1021 ttagtgctac tggttcttgt ggtttgaatt gcgacgcatc acttgaactt attattttcc
     1081 tttttcaccc aaatgtttta cctcgttgaa aatgtgccat ttacacccta gtgtgttttg
     1141 aactgaacta ttgagtaacg cgtgtctgtc cccaaataat acctgtaaag cgttgtattt
     1201 ctgtaaggat tctgcgttta gacctaactg tggttctgtt aaccagtctg agttgaccca
     1261 cgttttggat cgcttgttgt aacgaatgta caagagtgct tatgttgtat aaaatattca
     1321 tggcattttg tgccaggcaa aattagtcaa tacatttgat ttacatgatt aattcatgca
     1381 gacatttgaa atatggatga agtcccaaat cactagtgaa ttcgcggccg cctgcaggtc
     1441 gaccatatgg gagagctccc aacgcgttgg atgcatagct tgagtattct atagtgtcac
     1501 ctaaatagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1561 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1621 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1681 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1741 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1801 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1861 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1921 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1981 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2041 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2101 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2161 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2221 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2281 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2341 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2401 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2461 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2521 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2581 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2641 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2701 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2761 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2821 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2881 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2941 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3001 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3061 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3121 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3181 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3241 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3301 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3361 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3421 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3481 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3541 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3661 cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     3721 catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3781 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3841 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3901 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3961 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     4021 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4081 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4141 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcaggc
     4201 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4261 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4321 gttgtaaaac gacggccagt gaattg
//
