LOCUS       sequence-225488-        6104 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes proteolytically cleaved murine VEGF-D deltaN deltaC 
            form, which has signal peptide from murine IL3 gene to be used for 
            stimulation of lymphangiogenesis in mice in vivo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6104)
  TITLE     Alitalo expression plasmids
REFERENCE   2  (bases 1 to 6104)
  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"
COMMENT     Sequence Label: psubCMV-WPRE-IL3SP-mVEGF-D-dNdC
FEATURES             Location/Qualifiers
     source          1..6104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   27..156
                     /label=AAV2 ITR
     enhancer        234..537
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        538..741
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     692..712
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     738..762
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          879..975
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     regulatory      1054..1063
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1523..2111
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1576..1596)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1994..2005)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2126..2260
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2150..2169)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2204..2223
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2410..2539
                     /label=AAV2 ITR
     rep_origin      2910..3423
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2997..3016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3207..3228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3905..3924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4024..4046
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4084..4102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4170..4274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4275..4343,4344..5135)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4493..4512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5306..5894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5795..5814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcgttggc cgattcatta atgcagctgc gcgctcgctc gctcactgag gccgcccggg
       61 caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca
      121 gagagggagt ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat
      181 gctacttatc tacgtagcca tgctctagaa ttcgagcttg catgcctgca ggtcgttaca
      241 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      301 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      361 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      421 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      481 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      541 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      601 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      661 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      721 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
      781 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccggac tctagaggat
      841 ccggtactcg aggaactgaa aaaccagaaa gttaactggt aagtttagtc tttttgtctt
      901 ttatttcagg tcccggatcc ggtggtggtg caaatcaaag aactgctcct cagtggatgt
      961 tgcctttact tctaggcctg tacggaagtg ttacttctgc tctaaaagct gcggaattgt
     1021 acccgcggcc gctagctgca gcacgtgacg cgtgccacca tggttcttgc cagctctacc
     1081 accagcatcc acaccatgct gctcctgctc ctgatgctct tccacctggg actccaagct
     1141 tcaatctcgg cgcgtaattt tgcggcaact ttctatgaca ctgaaacact aaaagttata
     1201 gatgaagaat ggcagaggac ccaatgcagc cctagagaga catgcgtaga agtcgccagt
     1261 gagctgggga agacaaccaa cacattcttc aagcccccct gtgtaaatgt cttccggtgt
     1321 ggaggctgct gcaacgaaga gggtgtgatg tgtatgaaca caagcacctc ctacatctcc
     1381 aaacagctct ttgagatatc agtgcctctg acatcagtgc ccgagttagt gcctgttaaa
     1441 attgccaacc atacgggttg taagtgcttg cccacgggcc cccgccatcc ttactcaatt
     1501 atcagaagat gaacgcgtgc ataatcaacc tctggattac aaaatttgtg aaagattgac
     1561 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1621 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1681 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     1741 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     1801 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     1861 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     1921 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     1981 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2041 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2101 cgcctccccg catcgcggcc gcggggatcc agacatgata agatacattg atgagtttgg
     2161 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2221 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2281 ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc tagagtcgac
     2341 ctgcaggcat gcaagctcta gagcatggct acgtagataa gtagcatggc gggttaatca
     2401 ttaactacaa ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
     2461 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
     2521 tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2581 aacagttgcg cagcctgaat ggcgaatggc gattccgttg caatggctgg cggtaatatt
     2641 gttctggata ttaccagcaa ggccgatagt ttgagttctt ctactcaggc aagtgatgtt
     2701 attactaatc aaagaagtat tgcgacaacg gttaatttgc gtgatggaca gactctttta
     2761 ctcggtggcc tcactgatta taaaaacact tctcaggatt ctggcgtacc gttcctgtct
     2821 aaaatccctt taatcggcct cctgtttagc tcccgctctg attctaacga ggaaagcacg
     2881 ttatacgtgc tcgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     2941 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3001 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3061 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3121 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3181 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3241 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3301 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     3361 aatttaaata tttgcttata caatcttcct gtttttgggg cttttctgat tatcaaccgg
     3421 ggtacatatg attgacatgc tagttttacg attaccgttc atcgattctc ttgtttgctc
     3481 cagactctca ggcaatgacc tgatagcctt tgtagagacc tctcaaaaat agctaccctc
     3541 tccggcatga atttatcagc tagaacggtt gaatatcata ttgatggtga tttgactgtc
     3601 tccggccttt ctcacccgtt tgaatcttta cctacacatt actcaggcat tgcatttaaa
     3661 atatatgagg gttctaaaaa tttttatcct tgcgttgaaa taaaggcttc tcccgcaaaa
     3721 gtattacagg gtcataatgt ttttggtaca accgatttag ctttatgctc tgaggcttta
     3781 ttgcttaatt ttgctaattc tttgccttgc ctgtatgatt tattggatgt tggaatcgcc
     3841 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc
     3901 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     3961 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4021 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4081 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4141 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4201 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4261 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4321 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4381 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4441 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4501 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4561 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     4621 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4681 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4741 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4801 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     4861 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     4921 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4981 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5041 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5101 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6001 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     6061 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct cccc
//
