LOCUS       sequence-261113-        5621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mouse PDGFb and katushka fluorescent reporter. Construct 
            has inverted repeats to be used in Sleeping Beauty transposon 
            system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5621)
  TITLE     Plasmids for sleeping beauty system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5621)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pKT-mPDGFb-IRES-Katushka
FEATURES             Location/Qualifiers
     source          1..5621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1567..2139
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1590..2141
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1734..1751)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             2141..2845
                     /codon_start=1
                     /product="far-red-shifted derivative of red fluorescent 
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
                     /label=TurboFP635
                     /note="mammalian codon-optimized"
                     /translation="MVGEDSVLITENMHMKLYMEGTVNDHHFKCTSEGEGKPYEGTQTM
                     KIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERITTYEDGG
                     VLTATQDTSLQNGCLIYNVKINGVNFPSNGPVMQKKTLGWEASTEMLYPADSGLRGHSQ
                     MALKLVGGGYLHCSLKTTYRSKKPAKNLKMPGFYFVDRRLERIKEADKETYVEQHEMAV
                     ARYCDLPSKLGHS"
     polyA_signal    2924..3045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2935..2954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2989..3008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3147..3165
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3475..3492)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3646..4234)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3726..3745)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4377..5186)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5100..5119
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5178..5197
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5538..5557)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5539..5557)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gagtcaatgg gaaaaaccca ttggagccaa gtacactgac tcaataggga ctttccattg
       61 ggttttgccc agtacataag gtcaataggg ggtgagtcaa caggaaagtc ccattggagc
      121 caagtacatt gagtcaatag ggactttcca atgggttttg cccagtacat aaggtcaatg
      181 ggaggtaagc caatgggttt ttcccattac tgacatgtat actgagtcat tagggacttt
      241 ccaatgggtt ttgcccagta cataaggtca ataggggtga atcaacagga aagtcccatt
      301 ggagccaagt acactgagtc aatagggact ttccattggg ttttgcccag tacaaaaggt
      361 caataggggg tgagtcaatg ggtttttccc attattggca catacataag gtcaataggg
      421 gtgactagtg gagaagagca tgcttgaggg ctgagtgccc ctcagtgggc agagagcaca
      481 tggcccacag tccctgagaa gttgggggga ggggtgggca attgaactgg tgcctagaga
      541 aggtggggct tgggtaaact gggaaagtga tgtggtgtac tggctccacc tttttcccca
      601 gggtggggga gaaccatata taagtgcagt agtctctgtg aacattcaag cttctgcctt
      661 ctccctcctg tgagtttggt aagtcactga ctgtctatgc ctgggaaagg gtgggcagga
      721 gatggggcag tgcaggaaaa gtggcactat gaaccctgca gccctaggaa tgcatctagc
      781 atggccgcgg gattatgaat cgctgctggg cgctcttcct tcctctctgc tgctacctgc
      841 gcctggtcag cgccgagggg gatcccattc ctgaggaact gtatgaaatg ctgagcgacc
      901 actccatccg ctcctttgat gatcttcagc gcctgctgca cagagactcc gtagatgaag
      961 atggggctga gctggacttg aacatgaccc gagcacactc cggagtcgag ttggaaagct
     1021 catctcgagg gaggaggagc ctagggtccc tggcagcagc agagcctgct gtaatcgccg
     1081 agtgcaagac gcgcacagag gtgttccaga tctcgcggaa cctcatcgat cgcaccaacg
     1141 ccaacttcct ggtgtggccg ccctgtgtgg aggtgcagcg ctgctccggc tgctgcaata
     1201 accgcaatgt gcaatgccgg gcctcgcagg tgcagatgcg gccggtccag gtgagaaaga
     1261 ttgagattgt gcgaaagaag cccatcttca agaaggccac agtgaccttg gaggaccacc
     1321 tcgcctgcaa gtgtgagaca atagtgaccc ctcggcctgt gactagaagt cctgggacat
     1381 ccagggagca gcgagccaag acgcctcaag ctcgggtgac cattcggacg gtgagaatcc
     1441 gccggccccc caaaggcaag caccgaaagt ttaagcacac ccatgacaag gcggccctga
     1501 aggagaccct cggagcctag aatcactagt gcggccgcct gcaggtcgac catatgggag
     1561 tcgaggcccc tctccctccc ccccccctaa cgttactggc cgaagccgct tggaataagg
     1621 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
     1681 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
     1741 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
     1801 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
     1861 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
     1921 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctcacct caagcgtatt
     1981 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
     2041 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aacgtctagg ccccccgaac
     2101 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggtgggtg aggatagcgt
     2161 gctgatcacc gagaacatgc acatgaaact gtacatggag ggcaccgtga acgaccacca
     2221 cttcaagtgc acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgaagat
     2281 caaggtggtc gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat
     2341 gtacggcagc aaaaccttta tcaaccacac ccagggcatc cccgacttct ttaagcagtc
     2401 cttccctgag ggcttcacat gggagaggat caccacatac gaagacgggg gcgtgctgac
     2461 cgctacccag gacaccagcc tccagaacgg ctgcctcatc tacaacgtca agatcaacgg
     2521 ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccag
     2581 caccgagatg ctgtaccccg ctgacagcgg cctgagaggc catagccaga tggccctgaa
     2641 gctcgtgggc gggggctacc tgcactgctc cctcaagacc acatacagat ccaagaaacc
     2701 cgctaagaac ctcaagatgc ccggcttcta cttcgtggac aggagactgg aaagaatcaa
     2761 ggaggccgac aaagagacct acgtcgagca gcacgagatg gctgtggcca ggtactgcga
     2821 cctgcctagc aaactggggc acagctccgg actcagataa cgtacgtgcg cgcgaatgca
     2881 tccagatcta ggtaccaggg cccagctagc tggccagaca tgataagata cattgatgag
     2941 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3001 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3061 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3121 ctctacaaat gtggtatgga attatcaatt aaccctcact aaagggagac caagttaaac
     3181 aatttaaagg caatgctacc aaatactaat tgagtgtatg taaacttctg acccactggg
     3241 aatgtgatga aagaaataaa agctgaaatg aatcattctc tctactatta ttctgatatt
     3301 tcacattctt aaaataaagt ggtgatccta actgacctaa gacagggaat ttttactagg
     3361 attaaatgtc aggaattgtg aaaaagtgag tttaaatgta tttggctaag gtgtatgtaa
     3421 acttccgact tcaactgtat agggatctgg taccatttaa atctgttccg cttcctcgct
     3481 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3541 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3601 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3661 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3721 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3781 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaaccgtgg cgctttctca
     3841 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3901 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3961 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4021 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4081 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4141 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4201 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4261 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4321 aaggatcttc acctagatcc tttttgccag tgttacaacc aattaaccaa ttctgattag
     4381 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     4441 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     4501 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     4561 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     4621 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     4681 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     4741 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     4801 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     4861 tctaatacct ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca tgcatcatca
     4921 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     4981 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     5041 tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg cccgacatta
     5101 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     5161 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     5221 agttttattg ttcatgatgc agctggatcc agatccctat acagttgaag tcggaagttt
     5281 acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca aatttcttgt
     5341 taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg acacaagtca
     5401 tttttccaac aattgtttac agacagatta tttcacttat aattcactgt atcacaattc
     5461 cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc ttggaaaatt
     5521 ccagaaaatg atggtctccc tatagtgagt cgtattagat cccgggatta ctagtttaaa
     5581 cctaggatgc ataccggtca agctagcgat aattcctgca g
//
