LOCUS       sequence-308974-        6864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid for production of transgenic rainbow trout strain
            carrying the DsRed gene under control of the inhibin alpha 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6864)
  AUTHORS   Iwasaki-Takahashi Y, Shikina S, Watanabe M, Banba A, Yagisawa M, 
            Takahashi K, Fujihara R, Okabe T, Valdez DM Jr, Yamauchi A, 
            Yoshizaki G
  TITLE     Production of functional eggs and sperm from in vitro-expanded type 
            A spermatogonia in rainbow trout.
  JOURNAL   Commun Biol. 2020 Jun 15;3(1):308. doi: 10.1038/s42003-020-1025-y.
  PUBMED    32541813
REFERENCE   2  (bases 1 to 6864)
  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: "Commun 
            Biol."; date: "2020-06-15"; volume: "3(1):308. doi"; pages: " 
            10.1038/s42003-020-1025-y"
COMMENT     Sequence Label: inhibin-DsRed
FEATURES             Location/Qualifiers
     source          1..6864
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      2238..2247
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2244..2921
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2424..2444)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2836..2859
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3695..3816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3732..3751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3786..3805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3823..4278)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3960..3981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4172..4191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4305..4409
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4411..4768
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4411..4602
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4434..4454)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4619..4754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4803..5597
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4803..5597
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4857..4876)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5467..5486
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5785..5804)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5829..5876
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6205..6793
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6694..6713
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat tactagcgct
       61 accggactca gatctcgagc tcaagcttaa taagtatttg gtgaacaaca aacgtttatc
      121 tcaatacttt gttatatacc ctttgttggc aatgacagag gtcaaacgtt ttctgtaagt
      181 cttcacaagg ttttcacaca ctgttgctgg tattttggcc cattcctcca tgcagatctc
      241 ctctagagca gtgatgtttt ggtgctgttg ctgggcaaca cggactttca actccctcca
      301 aagattttct atggggttga gatctggaga ctggctaggc cgctcgagga ccttgaaatg
      361 cttcttacga agccactcct tcgttgcccg ggcggtgtgt ttgggatcat cgtcatgctg
      421 aaagacccag ccacgtttca tcttaaatgc ccttgttgat ggtaggcttt gttactttgg
      481 tccgagctct ctgcaggtca ttcactaggt ccccccgtgt ggttctggga tttttgctca
      541 ccgttcttgt gatcattttg accccacggg gtgagatctt gcgtggagcc ccagatcgag
      601 ggagattatc agtggtcttg tatgtcttcc atttcctaat aattgctccc acagttgatt
      661 tcttcaaacc aagctgctta cctattgcag attcagtctt cccagcctga tgcaggtcta
      721 caattttgtt tctggtgtcc tttgacagct ctttggtctt ggccatagtg gagtttggag
      781 tgtgactgtt tgaggttgtg gacaggtgta tttcatactg ataacaagtt caaacaggtg
      841 ccattaatac aggtaacgag tggaggacag agaagcctct taaagaagtt acaggtctgt
      901 gagagccaga aatcttgctt gtttgtaggt gaccaaatac ttattttccg acataatttg
      961 caaatacatt aattaaaaat gctacaatgt gattttctgg atttcttttc tcattttgtc
     1021 tgtcatagtt gaagtgtacc tatgatgaaa attacaggcc tctctcatcg ttttaagtgg
     1081 gagaacttgc acaattggtg gctggtagtt tttttgcgcc actgtacaca tatgttgggt
     1141 tcattatgat gaaaaataaa tcataatgaa cctttatttg atcagggagt cctactgaga
     1201 ccaaggtgtc atctacagat gagccttatg aaatgactac agcagcactg tgagttcacg
     1261 tgtgattact gttgtgatct attcgtaaca ctttatcttc taagggggtt gcaaagggtt
     1321 catgagtcgt ttatataaac agttataaca cattagttca aactgtggtt cattttcaat
     1381 aactgacccc aatatctgtt tacgtttgag tcattcaaca gacactccta tcaggagtga
     1441 cttacagtgc attcatctta agatagctag gtgagacaac cacattttct gcacttacag
     1501 agcctgtaaa ccgttccatt tcagacacct ctttcaatcg tgtcttatct ggccctggtg
     1561 ttttgtctaa tactcctact ctgctgaaat ctcctcacac ctccgacact aaatacacac
     1621 actatgtgtc ttaaagaaaa ccttaaacac gaaggaaaaa tacctcagaa atggtttcaa
     1681 agttttgcta cagtccactt cacaaaacag acaaatctag tacagtacaa gaacaaagtg
     1741 gcccttaccg tgaaacgcag actgtgatca aatacattgt aacttccaat cacccacaag
     1801 gtcatactct gaagtctttc ctctggataa tgactgcata acaataacat agggctctct
     1861 ctcactccct ttttctccag tgtctagata ggcatgggta ggaggcaata ttattccatg
     1921 acaaaacaat ggataaggga gacacagtgc tctgcgttgg cagagatggg aacctccaaa
     1981 taatacagga cttcattgac accactttcc cactggctca ggacagagaa ggatctacac
     2041 tacttcacct catctactag aagttttggc acaatcaatt gacccatttg gtcaccttac
     2101 caattattag acagggatat ctgacctact tggattactc aaactcaaag cacttttcca
     2161 cctcgatcta catgcttcac aaccctcccc tatagactga agagatctga accatgcaga
     2221 ctggggatcc accggtcgcc accatggcct cctccgagaa cgtcatcacc gagttcatgc
     2281 gcttcaaggt gcgcatggag ggcaccgtga acggccacga gttcgagatc gagggcgagg
     2341 gcgagggccg cccctacgag ggccacaaca ccgtgaagct gaaggtgacc aagggcggcc
     2401 ccctgccctt cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg
     2461 tgaagcaccc cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt
     2521 gggagcgcgt gatgaacttc gaggacggcg gcgtggcgac cgtgacccag gactcctccc
     2581 tgcaggacgg ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg
     2641 gccccgtgat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     2701 gcgacggcgt gctgaagggc gagacccaca aggccctgaa gctgaaggac ggcggccact
     2761 acctggtgga gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact
     2821 actacgtgga cgccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc
     2881 agtacgagcg caccgagggc cgccaccacc tgttcctgta gcggcctaaa cagactgtac
     2941 ccatcccaaa cgaccgaccc agccacaccc gactaccact tcagctctgc caccagacac
     3001 accgcccaga gggagagaga gagggggcct gcggagtgag cccaaaccca gcttctcagt
     3061 ctcattggca tggcttctgt tattttggcg cttttttttg actcaggatc tgtggaaaaa
     3121 caaaaaaaat atacacaact ggaacggtga aacagcggat taaagatttt taagagttaa
     3181 aaaaataaaa taatgtaaaa ggacccctga gattctatca tgcagctgag gactttaaaa
     3241 acaacaacat gtacattctc tttcttttcg agtcattcca attgttttgt taacttttca
     3301 gaacaggtat ttgcctctgt gaaggctgcc ggcgtccggc tctctgccac agcgttctgt
     3361 aacatggggc agtatggctt gtatgtaaat tatgtcataa tgtctgggtt tttttgtact
     3421 tgcagcctta agtcttggtc ctgtttaatt ttttttgttt ttgtttatgc aaatgaaccc
     3481 gagtgtgacc tctgctgggg ggggtcagag gtgattaggg tgccagaggg accagtgggg
     3541 ggggcaactt gtacactgac taaactccca ataaaggccg cgactctaga tcataatcag
     3601 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3661 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3721 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3781 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3841 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3901 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3961 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     4021 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     4081 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     4141 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     4201 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     4261 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     4321 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     4381 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     4441 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4501 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4561 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4621 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4681 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4741 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4801 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4861 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4921 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4981 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5041 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5101 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5161 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5221 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5281 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     5341 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5401 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5461 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5521 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5581 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5641 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5701 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5761 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5821 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5881 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5941 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     6001 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     6061 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6121 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6181 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6241 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6301 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6361 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6421 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6481 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6541 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6601 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6661 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6721 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6781 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6841 ttttgctcac atgttctttc ctgc
//
