LOCUS       sequence-231686-        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   300..429
                     /label=AAV2 ITR
     enhancer        507..810
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        811..1014
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     965..985
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1011..1035
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1152..1248
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     misc_feature    1760..2348
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1813..1833)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2231..2242)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2363..2497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2387..2406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2441..2460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2647..2776
                     /label=AAV2 ITR
     rep_origin      3147..3660
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3234..3253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3444..3465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4142..4161)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4261..4283
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4321..4339)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4407..4511
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4512..4580,4581..5372)
                     /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(4730..4749)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5543..6068,1..63)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6032..6051
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
       61 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
      121 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
      181 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
      241 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
      301 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
      361 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      421 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
      481 gaattcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
      541 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      601 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      661 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      721 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      781 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      841 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      901 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      961 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
     1021 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1081 cgggaccgat ccagcctccg gactctagag gatccggtac tcgaggaact gaaaaaccag
     1141 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
     1201 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
     1261 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg gccgctagct gcagcacgtg
     1321 acgcgtgcca ccatgtacgg cgagtggggc atgggcaaca tcctgatgat gttccacgtg
     1381 tacctggtgc agggcttcgc cgccaccttc tacgacaccg agaccctgaa ggtgatcgac
     1441 gaggaatggc agaggaccca gtgcagcccc agggaaacct gcgtggaggt ggccagcgag
     1501 ctgggcaaga ccaccaacac attcttcaag cccccctgcg tgaacgtgtt cagatgcggc
     1561 ggctgctgca acgaggaagg cgtgatgtgc atgaacacca gcaccagcta catcagcaag
     1621 cagctgttcg agatcagcgt gcccctgacc agcgtgcccg agctggtgcc cgtgaagatc
     1681 gccaaccaca ccggctgcaa gtgcctgccc accggcccca ggcaccccta cagcatcatc
     1741 agaaggtgaa cgcgtcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1801 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1861 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1921 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1981 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2041 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2101 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2161 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2221 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2281 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2341 ctccccgcat cgcggccgcg gggatccaga catgataaga tacattgatg agtttggaca
     2401 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2461 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2521 tatgtttcag gttcaggggg aggtgtggga ggttttttcg gatcctctag agtcgacctg
     2581 caggcatgca agctctagag catggctacg tagataagta gcatggcggg ttaatcatta
     2641 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2701 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     2761 gcgagcgagc gcgcagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     2821 agttgcgcag cctgaatggc gaatggcgat tccgttgcaa tggctggcgg taatattgtt
     2881 ctggatatta ccagcaaggc cgatagtttg agttcttcta ctcaggcaag tgatgttatt
     2941 actaatcaaa gaagtattgc gacaacggtt aatttgcgtg atggacagac tcttttactc
     3001 ggtggcctca ctgattataa aaacacttct caggattctg gcgtaccgtt cctgtctaaa
     3061 atccctttaa tcggcctcct gtttagctcc cgctctgatt ctaacgagga aagcacgtta
     3121 tacgtgctcg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3181 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3241 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3301 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3361 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3421 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3481 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3541 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     3601 ttaaatattt gcttatacaa tcttcctgtt tttggggctt ttctgattat caaccggggt
     3661 acatatgatt gacatgctag ttttacgatt accgttcatc gattctcttg tttgctccag
     3721 actctcaggc aatgacctga tagcctttgt agagacctct caaaaatagc taccctctcc
     3781 ggcatgaatt tatcagctag aacggttgaa tatcatattg atggtgattt gactgtctcc
     3841 ggcctttctc acccgtttga atctttacct acacattact caggcattgc atttaaaata
     3901 tatgagggtt ctaaaaattt ttatccttgc gttgaaataa aggcttctcc cgcaaaagta
     3961 ttacagggtc ataatgtttt tggtacaacc gatttagctt tatgctctga ggctttattg
     4021 cttaattttg ctaattcttt gccttgcctg tatgatttat tggatgttgg aatcgcctga
     4081 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
     4141 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4201 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4261 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4321 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4381 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4441 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     4501 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4561 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4621 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4681 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4741 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4801 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4861 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4921 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4981 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5041 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5101 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5161 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5221 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5281 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5341 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5401 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5461 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5521 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5581 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5641 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     5701 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5761 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5821 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5881 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5941 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6001 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6061 tcgggttt
//
