LOCUS       sequence-225485-        6738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes full-length (unprocessed) murine VEGF-D 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 6738)
  TITLE     Alitalo expression plasmids
REFERENCE   2  (bases 1 to 6738)
  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-WPRE
FEATURES             Location/Qualifiers
     source          1..6738
                     /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    2157..2745
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2210..2230)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2628..2639)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2760..2894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2784..2803)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2838..2857
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3044..3173
                     /label=AAV2 ITR
     rep_origin      3544..4057
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3631..3650)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3841..3862
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4539..4558)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4658..4680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4718..4736)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4804..4908
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4909..4977,4978..5769)
                     /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(5127..5146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5940..6528
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6429..6448
                     /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 gctagggccg ctccggatcc accatgtatg gagaatgggg aatggggaat
     1081 atcctcatga tgttccatgt gtacttggtg cagggcttca ggagcgaaca tggaccagtg
     1141 aaggattttt cttttgagcg atcatcccgg tccatgttgg aacgatctga acaacagatc
     1201 cgagcagctt ctagtttgga ggagttgctg caaatcgcgc actctgagga ctggaagctg
     1261 tggcgatgcc ggttgaagct caaaagtctt gccagtatgg actcacgctc agcatcccat
     1321 cgctccacca gatttgcggc aactttctat gacactgaaa cactaaaagt tatagatgaa
     1381 gaatggcaga ggacccaatg cagccctaga gagacatgcg tagaagtcgc cagtgagctg
     1441 gggaagacaa ccaacacatt cttcaagccc ccctgtgtaa atgtcttccg gtgtggaggc
     1501 tgctgcaacg aagagggtgt gatgtgtatg aacacaagca cctcctacat ctccaaacag
     1561 ctctttgaga tatcagtgcc tctgacatca gtgcccgagt tagtgcctgt taaaattgcc
     1621 aaccatacgg gttgtaagtg cttgcccacg ggcccccgcc atccttactc aattatcaga
     1681 agatccattc agaccccaga agaagatgaa tgtcctcatt ccaagaaact ctgtcctatt
     1741 gacatgctgt gggataacac caaatgtaaa tgtgttttgc aagacgagac tccactgcct
     1801 gggacagaag accactctta cctccaggaa cccactctct gtggaccgca catgacgttt
     1861 gatgaagatc gctgtgagtg cgtctgtaaa gcaccatgtc cgggagatct cattcagcac
     1921 ccggaaaact gcagttgctt tgagtgcaaa gaaagtctgg agagctgctg ccaaaagcac
     1981 aagatttttc acccagacac ctgcagctgt gaggacagat gtccttttca caccagaaca
     2041 tgtgcaagta gaaagccagc ctgtggaaag cactggcgct ttccaaagga gacaagggcc
     2101 cagggactct acagccagga gaacccttga gctagctccg gagcggccgc gtcgataatc
     2161 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     2221 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     2281 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     2341 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     2401 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     2461 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     2521 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     2581 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     2641 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     2701 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcgc ggccgcgggg
     2761 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2821 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2881 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2941 tgtgggaggt tttttcggat cctctagagt cgacctgcag gcatgcaagc tctagagcat
     3001 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     3061 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3121 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctggcgt
     3181 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3241 tggcgattcc gttgcaatgg ctggcggtaa tattgttctg gatattacca gcaaggccga
     3301 tagtttgagt tcttctactc aggcaagtga tgttattact aatcaaagaa gtattgcgac
     3361 aacggttaat ttgcgtgatg gacagactct tttactcggt ggcctcactg attataaaaa
     3421 cacttctcag gattctggcg taccgttcct gtctaaaatc cctttaatcg gcctcctgtt
     3481 tagctcccgc tctgattcta acgaggaaag cacgttatac gtgctcgtca aagcaaccat
     3541 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3601 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3661 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3721 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3781 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3841 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3901 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3961 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta aatatttgct tatacaatct
     4021 tcctgttttt ggggcttttc tgattatcaa ccggggtaca tatgattgac atgctagttt
     4081 tacgattacc gttcatcgat tctcttgttt gctccagact ctcaggcaat gacctgatag
     4141 cctttgtaga gacctctcaa aaatagctac cctctccggc atgaatttat cagctagaac
     4201 ggttgaatat catattgatg gtgatttgac tgtctccggc ctttctcacc cgtttgaatc
     4261 tttacctaca cattactcag gcattgcatt taaaatatat gagggttcta aaaattttta
     4321 tccttgcgtt gaaataaagg cttctcccgc aaaagtatta cagggtcata atgtttttgg
     4381 tacaaccgat ttagctttat gctctgaggc tttattgctt aattttgcta attctttgcc
     4441 ttgcctgtat gatttattgg atgttggaat cgcctgatgc ggtattttct ccttacgcat
     4501 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     4561 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4621 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4681 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     4741 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     4801 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4861 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4921 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4981 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5041 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5101 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5161 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5221 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5281 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5341 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5401 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5461 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5521 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5581 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5641 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5701 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5761 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5821 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5881 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5941 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6001 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6061 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6121 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6181 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6241 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6301 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6361 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6421 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6481 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6541 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6601 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6661 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     6721 cgcaaaccgc ctctcccc
//
