LOCUS       sequence-225487-        6068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes proteolytically cleaved murine VEGF-D deltaN deltaC 
            form, which has native signal peptide 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 6068)
  TITLE     Alitalo expression plasmids
REFERENCE   2  (bases 1 to 6068)
  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-mVEGF-D-dNdC-WPRE
FEATURES             Location/Qualifiers
     source          1..6068
                     /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"
     misc_feature    1487..2075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1540..1560)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1958..1969)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2090..2224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2114..2133)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2168..2187
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2374..2503
                     /label=AAV2 ITR
     rep_origin      2874..3387
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2961..2980)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3171..3192
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3869..3888)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3988..4010
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4048..4066)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4134..4238
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4239..4307,4308..5099)
                     /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(4457..4476)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5270..5858
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5759..5778
                     /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 tgtacggcga gtggggcatg
     1081 ggcaacatcc tgatgatgtt ccacgtgtac ctggtgcagg gcttcgccgc caccttctac
     1141 gacaccgaga ccctgaaggt gatcgacgag gaatggcaga ggacccagtg cagccccagg
     1201 gaaacctgcg tggaggtggc cagcgagctg ggcaagacca ccaacacatt cttcaagccc
     1261 ccctgcgtga acgtgttcag atgcggcggc tgctgcaacg aggaaggcgt gatgtgcatg
     1321 aacaccagca ccagctacat cagcaagcag ctgttcgaga tcagcgtgcc cctgaccagc
     1381 gtgcccgagc tggtgcccgt gaagatcgcc aaccacaccg gctgcaagtg cctgcccacc
     1441 ggccccaggc acccctacag catcatcaga aggtgaacgc gtgcataatc aacctctgga
     1501 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     1561 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     1621 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     1681 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     1741 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     1801 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     1861 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     1921 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     1981 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     2041 gacgagtcgg atctcccttt gggccgcctc cccgcatcgc ggccgcgggg atccagacat
     2101 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     2161 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2221 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2281 tttttcggat cctctagagt cgacctgcag gcatgcaagc tctagagcat ggctacgtag
     2341 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     2401 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     2461 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt aatagcgaag
     2521 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgattcc
     2581 gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga tagtttgagt
     2641 tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgcgac aacggttaat
     2701 ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa cacttctcag
     2761 gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt tagctcccgc
     2821 tctgattcta acgaggaaag cacgttatac gtgctcgtca aagcaaccat agtacgcgcc
     2881 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2941 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3001 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3061 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3121 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3181 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3241 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3301 ttttaacaaa atattaacgc ttacaattta aatatttgct tatacaatct tcctgttttt
     3361 ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt tacgattacc
     3421 gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag cctttgtaga
     3481 gacctctcaa aaatagctac cctctccggc atgaatttat cagctagaac ggttgaatat
     3541 catattgatg gtgatttgac tgtctccggc ctttctcacc cgtttgaatc tttacctaca
     3601 cattactcag gcattgcatt taaaatatat gagggttcta aaaattttta tccttgcgtt
     3661 gaaataaagg cttctcccgc aaaagtatta cagggtcata atgtttttgg tacaaccgat
     3721 ttagctttat gctctgaggc tttattgctt aattttgcta attctttgcc ttgcctgtat
     3781 gatttattgg atgttggaat cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3841 tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3901 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3961 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4021 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4081 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4141 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4201 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4261 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4321 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4381 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4441 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4501 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4561 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4621 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4681 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4741 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4801 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4861 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4921 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4981 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5041 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5101 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5161 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5221 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5281 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5341 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5401 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5461 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5521 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5581 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5641 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5701 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5761 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5821 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5881 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     5941 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6001 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     6061 ctctcccc
//
