LOCUS       sequence-205155-        5783 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid can be used for secretion of fish Interferon gamma.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5783)
  AUTHORS   Zahradnik J, Kolarova L, Parizkova H, Kolenko P, Schneider B
  TITLE     Interferons type II and their receptors R1 and R2 in fish species: 
            Evolution, structure, and function.
  JOURNAL   Fish Shellfish Immunol. 2018 May 6;79:140-152. doi: 
            10.1016/j.fsi.2018.05.008.
  PUBMED    29742458
REFERENCE   2  (bases 1 to 5783)
  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: "Fish 
            Shellfish Immunol."; date: "2018-05-6"; volume: "79:140-152. doi"; 
            pages: " 10.1016/j.fsi.2018.05.008"
COMMENT     Sequence Label: OnilIFN-gamma
FEATURES             Location/Qualifiers
     source          1..5783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             210..232
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             804..821
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(874..892)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      888..935
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      972..1427
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1059..1078)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1269..1290
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1519..2334)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1519..2334)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2242..2261
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2326..2345
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2456..3044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2945..2964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3198..3215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3230..3369
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3295..3314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3455..3477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3471..3662)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4471..5553)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5515..5534
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5554..5631)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
       61 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      121 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggtgtga
      181 gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata
      241 tgtcccacat cccagcagag atgaacttga ccatccatag gctgttgcag cactataaaa
      301 tttcaaacca ggaaagattt aatgggaaac ccgtatttcc tagtgaccag aacaaaaaaa
      361 tcgagaccaa gttgatcttc atgggtggtg ttttggagtc gtacgaaaag ctgcttggcc
      421 acatgttgaa acaactgccc actccgagtc cccagatcac cgggagcaac gcgcaacctg
      481 gctcggccat caccgccgcc tccagcagca ccaacaactc aggctcgcta ccaggcacag
      541 atgtcagatc agagctgaat tacatcctga agaaggtcca ggagctgaag aaacaccgct
      601 accatgagca ggagaaactt ctgcagggat tggagaccct cagagatatc aagatgaatg
      661 accacatcgt tcagagcaaa gcactttggc agctgccaca gctgtacgaa gaggcgagct
      721 cactccccga aatcaagatc cagaggagac gcagacggcg gaggcaaaca aaggctaaag
      781 gtcgccccag agtttgactc gagcaccacc accaccacca ctgagatccg gctgctaaca
      841 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      901 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
      961 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1021 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1081 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1141 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1201 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1261 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1321 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1381 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     1441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     1501 ccgctcatga attaattctt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     1561 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     1621 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     1681 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     1741 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     1801 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     1861 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     1921 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     1981 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
     2041 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2101 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2161 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
     2221 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     2281 gttggaattt aatcgcggcc tagagcaaga cgtttcccgt tgaatatggc tcataacacc
     2341 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgaccaaa atcccttaac
     2401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2581 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     2641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3061 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3121 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3181 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     3241 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     3301 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3361 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3421 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3481 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     3541 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     3601 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     3661 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     3721 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     3781 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     3841 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     3901 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3961 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     4021 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     4081 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     4141 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     4201 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     4261 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     4321 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4381 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     4441 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4501 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4561 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     4621 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     4681 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     4741 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     4801 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     4861 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     4921 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4981 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     5041 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     5101 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     5161 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     5221 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     5281 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     5341 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5401 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     5461 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     5521 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     5581 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     5641 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     5701 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     5761 ccacggggcc tgccaccata ccc
//
