LOCUS       sequence-408672-        7418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A donor vector plasmid for Cre-mediated integration of paromomycin 
            resistance (Paro) and dog IFN-alpha-4 (IFN-alpha-4) genes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7418)
  AUTHORS   Huang G, Kawabe Y, Shirakawa K, Akiyama T, Kamihira M
  TITLE     Novel transgenic Chlamydomonas reinhardtii strain with retargetable 
            genomic transgene integration using Cre-loxP system.
  JOURNAL   J Biosci Bioeng. 2021 Nov;132(5):469-478. doi: 
            10.1016/j.jbiosc.2021.07.006. Epub 2021 Aug 19.
  PUBMED    34420898
REFERENCE   2  (bases 1 to 7418)
  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: 
            "10.1016/j.jbiosc.2021.07.006"; journalName: "J Biosci Bioeng"; 
            date: "2021-11"; volume: "132"; issue: "5"; pages: "469-478"
COMMENT     Sequence Label: pD/Paro-IFN
FEATURES             Location/Qualifiers
     source          1..7418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(785..802)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    823..856
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             complement(4283..5077)
                     /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             complement(4283..5077)
                     /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(4394..4413)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5004..5023
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5441..5458)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      5611..5697
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5789..5816
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      5950..6538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6439..6458
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6692..6709
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6724..6864
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6790..6809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6950..6972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tcttctctca tccgccaaaa cagccaagct ggagaccgtt tgacattacc ctgttatccc
       61 tagatacatt accctgttat cccagatgac ataccctgtt atccctagat gacattaccc
      121 tgttatccca gatgacatta ccctgttatc cctagataca ttaccctgtt atcccagatg
      181 acataccctg ttatccctag atgacattac cctgttatcc cagatgacat taccctgtta
      241 tccctagata cattaccctg ttatcccaga tgacataccc tgttatccct agatgacatt
      301 accctgttat cccagatgac attaccctgt tatccctaga tacattaccc tgttatccca
      361 gatgacatac cctgttatcc ctagatgaca ttaccctgtt atcccagatg acattaccct
      421 gttatcccta gatacattac cctgttatcc cagatgacat accctgttat ccctagatga
      481 cattaccctg ttatcccaga tgacattacc ctgttatccc tagatacatt accctgttat
      541 cccagatgac ataccctgtt atccctagat gacattaccc tgttatccca gatgacatta
      601 ccctgttatc cctagataca ttaccctgtt atcccagatg acataccctg ttatccctag
      661 atgacattac cctgttatcc cagatgacat taccctgtta tccctagata cattaccctg
      721 ttatcccaga tgacataccc tgttatccct agatgacatt accctgttat cccagataaa
      781 ctcaatgatg atgatgatga tggtcgagac tcagcggccg cggtgccagg gcgtgccctt
      841 gggctccccg ggcgcgacta gaggatccat aacttcgtat aaccataatt atacgaacgg
      901 taatcgatgc tagtgacggc ggggagctcg ctgaggcttg acatgattgg tgcgtatgtt
      961 tgtatgaagc tacaggactg atttggcggg ctatgagggc gggggaagct ctggaagggc
     1021 cgcgatgggg cgcgcggcgt ccagaaggcg ccatacggcc cgctggcggc acccatccgg
     1081 tataaaagcc cgcgaccccg aacggtgacc tccactttca gcgacaaacg agcacttata
     1141 catacgcgac tattctgccg ctatacataa ccactcaagc ttgcatgccg ggcgcgccag
     1201 aaggagcgca gccaaaccag gatgatgttt gatggggtat ttgagcactt gcaaccctta
     1261 tccggaagcc ccctggccca caaaggctag gcgccaatgc aagcagttcg catgcagccc
     1321 ctggagcggt gccctcctga taaaccggcc agggggccta tgttctttac ttttttacaa
     1381 gagaagtcac tcaacatctt aaaatggcca ggtgagtcga cgagcaagcc cggcggatca
     1441 ggcagcgtgc ttgcagattt gacttgcaac gcccgcattg tgtcgacgaa ggcttttggc
     1501 tcctctgtcg ctgtctcaag cagcatctaa ccctgcgtcg ccgtttccat ttgcaggatg
     1561 catatgtctc gctcccccag catcgcaatt gtcctcgcgg tgagtcgacg agcaagcccg
     1621 gcggatcagg cagcgtgctt gcagatttga cttgcaacgc ccgcattgtg tcgacgaagg
     1681 cttttggctc ctctgtcgct gtctcaagca gcatctaacc ctgcgtcgcc gtttccattt
     1741 gcaggcggtc gccctcctgg gcgtctgcgc gctggcccag agatctgacg tctgtgatct
     1801 gcccgacacc cactctttac gcaactggcg cgtgctgact ttactggggc aaatgcgccg
     1861 tttaagcgcc agctcttgtg accactacac caccgacttc gccttcccca aggagctgtt
     1921 cgacggccag cgtttacaag aggcccaagc cttaagcgtg gtgcacgtga tgacccagaa
     1981 ggtgttccat ttattctgca ccaacatgag cagcgcccct tggaacatga ctttactgga
     2041 ggagctgtgc tcggtgagct tgcggggttg cgagcaacac tccagcaacg aacagtgccc
     2101 aagtcaggaa tctgcagtca gcctgggctt tcggcggctt tttcttgggc aaacagcttg
     2161 cactcatgcc agcgcggctt gtccagcctc acttgagctt tccagctgct accagccggg
     2221 ctatacgaca gcgacagagc catagcgtgg aatcacttat ttgggttgcc gaagtagcgg
     2281 tcggagcgtg agttcttggt caagccgccc cttatccggt tcctgtccgt gtctttgtcc
     2341 ctcgttcacc cttcgcggca cccttcatcc ccttgcttgc agggtttaag cgagcagctg
     2401 gacgatttag acgcttgtcc tttacaagaa gccggtttag ccgagacccc tttaatgcac
     2461 gaggacagca ctttacgcac ctacttccag cgcatctctt tatatttaca agatcgcaac
     2521 cacagccctt gtgcttggga aatggtgcgc gcggagatcg gccgcagctt cttctcttta
     2581 accattttac aagagcgtgt tcgccgccgc aagtgaacgc gtccgctccg tgtaaatgga
     2641 ggcgctcgtt gatctgagcc ttgccccctg acgaacggcg gtggatggaa gatactgctc
     2701 tcaagtgctg aagcggtagc ttagctcccc gtttcgtgct gatcagtctt tttcaacacg
     2761 taaaaagcgg aggagttttg caattttgtt ggttgtaacg atcctccgtt gattttggcc
     2821 tctttctcca tgggcgggct gggcgtattt gaagcggacc cggatcctac cgttcgtata
     2881 aagtatccta tacgaagtta taggcagcgt gcttgcagat ttgacttgca acgcccgcat
     2941 tgtgtcgacg aaggcttttg gctcctctgt cgctgtctca agcagcatct aaccctgcgt
     3001 cgccgtttcc atttgcaggt ttcgaagcat ggacgatgcg ttgcgtgcac tgcggggtcg
     3061 gtatcccggt tgtgagtggg ttgttgtgga ggatggggcc tcgggggctg gtgtttatcg
     3121 gcttcggggt ggtgggcggg agttgtttgt caaggtggca gctctggggg ccggggtggg
     3181 cttgttgggt gaggctgagc ggctggtgtg gttggcggag gtggggattc ccgtacctcg
     3241 tgttgtggag ggtggtgggg acgagagggt cgcctggttg gtcaccgaag cggttccggg
     3301 gcgtccggcc agtgcgcggt ggccgcggga gcagcggctg gacgtggcgg tggcgctcgc
     3361 ggggctcgct cgttcgctgc acgcgctgga ctgggagcgg tgtccgttcg atcgcagtct
     3421 cgcggtgacg gtgccgcagg cggcccgtgc tgtcgctgaa gggagcgtcg acttggagga
     3481 tctggacgag gagcggaagg ggtggtcggg ggagcggctt ctcgccgagc tggagcggac
     3541 tcggcctgcg gacgaggatc tggcggtttg ccacggtgac ctgtgcccgg acaacgtgct
     3601 gctcgaccct cgtacctgcg aggtgaccgg gctgatcgac gtggggcggg tcggccgtgc
     3661 ggaccggcac tccgatctcg cgctggtgct gcgcgagctg gcccacgagg aggacccgtg
     3721 gttcgggccg gagtgttccg cggcgttcct gcgggagtac gggcgcgggt gggatggggc
     3781 ggtatcggag gaaaagctgg cgttttaccg gctgttggac gagttcttct gactcgagtg
     3841 accgctccgt gtaaatggag gcgctcgttg atctgagcct tgccccctga cgaacggcgg
     3901 tggatggaag atactgctct caagtgctga agcggtagct tagctccccg tttcgtgctg
     3961 atcagtcttt ttcaacacgt aaaaagcgga ggagttttgc aattttgttg gttgtaacga
     4021 tcctccgttg attttggcct ctttctccat gggcgggctg ggcgtatttg aagcggaccc
     4081 ggtaccacgc gtagatctgg taccataact tcgtataacc ataattatac gaagttatgt
     4141 cgacccatgg gggcccgccc caactggggt aacctttgag ttctctcagt tgggggtaat
     4201 cagcatcatg atgtggtacc acatcatgat gctgattata agaatgcggc cgccacactc
     4261 tagtggatct cgagttaata attcagaaga actcgtcaag aaggcgatag aaggcgatgc
     4321 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     4381 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     4441 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     4501 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     4561 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     4621 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4681 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4741 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4801 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4861 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4921 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     4981 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     5041 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     5101 cttgatcaga gcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     5161 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     5221 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     5281 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     5341 ggggtcagca ccgtttctgc ggactggctt tctacgcgct cgaggggggg ccaaacggtc
     5401 tccagcttgg ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag
     5461 aacgcagaag cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac
     5521 ctgaccccat gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc
     5581 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     5641 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     5701 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     5761 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
     5821 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgac
     5881 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5941 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6001 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6061 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     6121 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6181 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6241 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6301 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6361 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6421 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6481 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6661 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6721 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
     6781 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     6841 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     6901 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     6961 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     7021 gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa
     7081 accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta tgacaacttg
     7141 acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg gctggccccg
     7201 gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac attgcgaccg
     7261 acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct gatacgttgg
     7321 tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg tgacagacgc
     7381 gacggcgaca agcaaacatg ctgtgcgacg ctggcgat
//
