LOCUS       sequence-205175-        5761 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 5761)
  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 5761)
  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: PoliIFN-gamma
FEATURES             Location/Qualifiers
     source          1..5761
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     160..179
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        160..178
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    179..203
                     /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)."
     RBS             218..240
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             659..670
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             785..802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(855..873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      869..916
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      953..1408
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1040..1059)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1250..1271
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1500..2315)
                     /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(1500..2315)
                     /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     2223..2242
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2307..2326
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2437..3025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2926..2945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3179..3196
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3211..3350
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3276..3295)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3436..3458
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3452..3643)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4452..5534)
                     /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     5496..5515
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5535..5612)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
       61 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
      121 cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg
      181 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
      241 tatacatatg tcacacatcc ctgcaaggat gaacaaaacc atccagaacc tcctgcagca
      301 ctataatatt tcaaacaaag acagattcaa cgggaagccc gtcttcccca aggagccgct
      361 gtccggcagg atggagacaa agatgctgtt catgggcggc gttttggaga cgtatgaaaa
      421 gctgataggc cagatgttgg agcagctgcc gaacacgact ccccccacag ccgggagcag
      481 agagggcctc aactccgctg ccccggaggt cagcgtcagg acggacctga actacattct
      541 gaagaaagtc caggagctga ggacaaatcg cttcaaggag cagagcaaac tgctgcaggg
      601 gctgcacgac ctcggagaca tcaagatgaa taacttcatc atccagagca aagcactgtg
      661 ggagctgcag tggatgtacg aggaggcgag ttctctgtca aacaacacca agatgcagag
      721 gaggcggcgg cggcggcgga gacaagctcg gaaggtcaaa actcctacga gagcctgact
      781 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
      841 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      901 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc
      961 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1021 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1081 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1141 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     1201 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     1261 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     1321 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     1381 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     1441 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg aattaattct
     1501 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     1561 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     1621 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     1681 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     1741 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     1801 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     1861 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     1921 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     1981 tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca
     2041 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     2101 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     2161 aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca
     2221 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     2281 ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg
     2341 taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2581 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3121 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3181 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3241 gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3301 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
     3361 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3421 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3481 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     3541 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     3601 taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag
     3661 ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac
     3721 gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg
     3781 tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata
     3841 cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa
     3901 tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
     3961 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
     4021 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
     4081 acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata atggcctgct
     4141 tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg gcgtgcaaga
     4201 ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag cggtcctcgc
     4261 cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     4321 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     4381 aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa cttacattaa
     4441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt
     4561 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc
     4621 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttaac
     4681 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca
     4741 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg
     4801 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa
     4861 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg
     4921 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct
     4981 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg
     5041 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac
     5101 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag
     5161 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct
     5221 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga
     5281 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg
     5341 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc
     5401 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg
     5461 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac
     5521 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata
     5581 ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc ccttatgcga
     5641 ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag
     5701 gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     5761 a
//
