LOCUS       sequence-358741-        7532 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Stably express PalmGamillus in mammalian cells via the sleeping 
            beauty transposon system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7532)
  AUTHORS   Perez GI, Broadbent D, Zarea AA, Dolgikh B, Bernard MP, Withrow A, 
            McGill A, Toomajian V, Thampy LK, Harkema J, Walker JR, Kirkland TA,
            Bachmann MH, Schmidt J, Kanada M
  TITLE     In Vitro and In Vivo Analysis of Extracellular Vesicle-Mediated 
            Metastasis Using a Bright, Red-Shifted Bioluminescent Reporter 
            Protein.
  JOURNAL   Adv Genet (Hoboken). 2022 Jan 19;3(1):2100055. doi: 
            10.1002/ggn2.202100055. eCollection 2022 Mar.
  PUBMED    36619349
REFERENCE   2  (bases 1 to 7532)
  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: "Adv Genet 
            (Hoboken)."; date: "2022-01-19"; volume: "3(1):2100055. doi"; pages:
            " 10.1002/ggn2.202100055. eCollection 2022 Mar"
COMMENT     Sequence Label: pKT2/CAGXSP/PalmGamillus
FEATURES             Location/Qualifiers
     source          1..7532
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          95..1111
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1035..1057
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1119..1138
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1374..2087
                     /codon_start=1
                     /product="acid-tolerant monomeric green fluorescent protein
                     (Shinoda et al., 2018)"
                     /label=Gamillus
                     /translation="VSKGEEASGRALFQYPMTSKIELNGEINGKKFKVAGEGFTPSSGR
                     FNMHAYCTTGDLPMSWVVIASPLQYGFHMFAHYPEDITHFFQECFPGSYTLDRTLRMEG
                     DGTLTTHHEYSLEDGCVTSKTTLNASGFDPKGATMTKSFVKQLPNEVKITPHGPNGIRL
                     TSTVLYLKEDGTIQIGTQDCIVTPVGGRKVTQPKAHFLHTQIIQKKDPNDTRDHIVQTE
                     LAVAGNLWHGMDELYK"
     primer_bind     complement(2148..2165)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2154..2378
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2424..2852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2511..2530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2721..2742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2861..2881)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2866..3195
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3046..3181
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3108..3127
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3206..3805
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3206..3225)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3702..3722
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3965..4086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4117..4135
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4445..4462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4616..5204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4696..4715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5347..6156)
                     /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     6070..6089
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6148..6167
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     polyA_signal    6661..6782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6845..6864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7036..7415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
       61 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      121 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      181 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta
      241 atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc
      301 tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc
      361 gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttgtgc
      421 gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg
      481 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
      541 gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg
      601 cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg
      661 ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggrgggctc
      721 gggggrgggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
      781 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
      841 tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg
      901 gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg
      961 tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg
     1021 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
     1081 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
     1141 gtctcatcat tttggcaaag aattcctcga ggaattcact cctcaggtgc aggctgccta
     1201 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctgca
     1261 tgagctagcg tttaaactta agcttggtac cgagctcgga tccactagtc cagtgtggtg
     1321 gaattctgca gatagccgcc accatgctgt gctgtatgag aagaaccaaa caggtgagca
     1381 agggcgagga ggcatctggc agagccctgt tccagtaccc catgaccagc aagatcgagc
     1441 tgaacggcga gatcaacggc aagaaattca aggtggccgg cgagggcttc acccccagca
     1501 gcggcagatt caacatgcac gcctactgca ccaccggcga cctgcctatg agctgggtcg
     1561 tgattgccag ccccctccag tacggcttcc acatgttcgc ccactacccc gaggacatca
     1621 cacacttttt ccaggaatgc ttccccggca gctacaccct ggaccggacc ctgagaatgg
     1681 aaggcgacgg caccctgacc acccaccacg agtacagcct ggaggacggc tgcgtgacct
     1741 ccaagaccac cctgaatgcc agcggcttcg accctaaggg cgccaccatg accaagagct
     1801 tcgtgaaaca actgcctaac gaggtgaaga tcacccccca cggccccaac ggcatcagac
     1861 tgaccagcac cgtgctgtac ctgaaggagg atggcaccat ccagatcggc acccaggact
     1921 gcatcgtgac ccctgtgggc ggaaggaaag tgacccagcc caaggcccac ttcctgcaca
     1981 cccagatcat ccagaagaag gaccccaacg acacccggga ccacatcgtg cagacagaac
     2041 tggccgtggc cggcaatctg tggcacggca tggacgagct gtacaagtaa atccagcaca
     2101 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     2161 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2221 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2281 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2341 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     2401 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2701 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2761 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2821 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     2881 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     2941 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3001 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3061 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3121 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3181 ctaggctttt gcaaaaagct cccgcatgac cgagtacaag cccacggtgc gcctcgccac
     3241 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3301 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3361 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3421 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3481 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3541 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     3601 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     3661 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     3721 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     3781 catgacccgc aagcccggtg cctgattcga aatgaccgac caagcgacgc ccaacctgcc
     3841 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4081 atcttatcat gtctgtatac cgtcgatagc gatatcaatt aaccctcact aaagggagac
     4141 caagttaaac aatttaaagg caatgctacc aaatactaat tgagtgtatg taaacttctg
     4201 acccactggg aatgtgatga aagaaataaa agctgaaatg aatcattctc tctactatta
     4261 ttctgatatt tcacattctt aaaataaagt ggtgatccta actgacctaa gacagggaat
     4321 ttttactagg attaaatgtc aggaattgtg aaaaagtgag tttaaatgta tttggctaag
     4381 gtgtatgtaa acttccgact tcaactgtat agggatctgg taccatttaa atctgttccg
     4441 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4501 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4561 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4621 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4681 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4741 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaaccgtgg
     4801 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4861 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4921 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4981 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5041 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5101 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5161 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5221 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5281 gattatcaaa aaggatcttc acctagatcc tttttgccag tgttacaacc aattaaccaa
     5341 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     5401 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     5461 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     5521 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     5581 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     5641 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     5701 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     5761 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     5821 aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg tgagtaacca
     5881 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     5941 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     6001 cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg cacctgattg
     6061 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     6121 tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6181 gtaagcagac agttttattg ttcatgatgc agctggatcc agatccctat acagttgaag
     6241 tcggaagttt acatacactt aagttggagt cattaaaact cgtttttcaa ctactccaca
     6301 aatttcttgt taacaaacaa tagttttggc aagtcagtta ggacatctac tttgtgcatg
     6361 acacaagtca tttttccaac aattgtttac agacagatta tttcacttat aattcactgt
     6421 atcacaattc cagtgggtca gaagtttaca tacactaagt tgactgtgcc tttaaacagc
     6481 ttggaagctg cgcactaggc aagttaacta actcctctga atgtcagtat ttccatctgt
     6541 aagatgaaca cagtggggct ccaattccat accacatttg tagaggtttt acttgcttta
     6601 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     6661 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6721 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6781 tatcatgtct ggccagctat cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6841 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     6901 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     6961 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     7021 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     7081 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7141 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7201 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7261 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     7321 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     7381 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     7441 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     7501 tattttgtgg gggcgaggcg gagaggtgcg gc
//
