LOCUS       sequence-231680-        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   312..441
                     /label=AAV2 ITR
     enhancer        519..822
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        823..1026
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     977..997
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1023..1047
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1164..1260
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     misc_feature    2442..3030
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2495..2515)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2913..2924)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3045..3179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3069..3088)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3123..3142
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3329..3458
                     /label=AAV2 ITR
     rep_origin      3829..4342
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3916..3935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4126..4147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4824..4843)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4943..4965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5003..5021)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5089..5193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5194..5262,5263..6054)
                     /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(5412..5431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6225..6738,1..75)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6714..6733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
       61 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      121 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      181 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      241 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
      301 cattaatgca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
      361 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      421 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      481 agccatgctc tagaattcga gcttgcatgc ctgcaggtcg ttacataact tacggtaaat
      541 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      601 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      661 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      721 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      781 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      841 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      901 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      961 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1021 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     1081 catagaagac accgggaccg atccagcctc cggactctag aggatccggt actcgaggaa
     1141 ctgaaaaacc agaaagttaa ctggtaagtt tagtcttttt gtcttttatt tcaggtcccg
     1201 gatccggtgg tggtgcaaat caaagaactg ctcctcagtg gatgttgcct ttacttctag
     1261 gcctgtacgg aagtgttact tctgctctaa aagctgcgga attgtacccg cggccgctag
     1321 ggccgctccg gatccaccat gtatggagaa tggggaatgg ggaatatcct catgatgttc
     1381 catgtgtact tggtgcaggg cttcaggagc gaacatggac cagtgaagga tttttctttt
     1441 gagcgatcat cccggtccat gttggaacga tctgaacaac agatccgagc agcttctagt
     1501 ttggaggagt tgctgcaaat cgcgcactct gaggactgga agctgtggcg atgccggttg
     1561 aagctcaaaa gtcttgccag tatggactca cgctcagcat cccatcgctc caccagattt
     1621 gcggcaactt tctatgacac tgaaacacta aaagttatag atgaagaatg gcagaggacc
     1681 caatgcagcc ctagagagac atgcgtagaa gtcgccagtg agctggggaa gacaaccaac
     1741 acattcttca agcccccctg tgtaaatgtc ttccggtgtg gaggctgctg caacgaagag
     1801 ggtgtgatgt gtatgaacac aagcacctcc tacatctcca aacagctctt tgagatatca
     1861 gtgcctctga catcagtgcc cgagttagtg cctgttaaaa ttgccaacca tacgggttgt
     1921 aagtgcttgc ccacgggccc ccgccatcct tactcaatta tcagaagatc cattcagacc
     1981 ccagaagaag atgaatgtcc tcattccaag aaactctgtc ctattgacat gctgtgggat
     2041 aacaccaaat gtaaatgtgt tttgcaagac gagactccac tgcctgggac agaagaccac
     2101 tcttacctcc aggaacccac tctctgtgga ccgcacatga cgtttgatga agatcgctgt
     2161 gagtgcgtct gtaaagcacc atgtccggga gatctcattc agcacccgga aaactgcagt
     2221 tgctttgagt gcaaagaaag tctggagagc tgctgccaaa agcacaagat ttttcaccca
     2281 gacacctgca gctgtgagga cagatgtcct tttcacacca gaacatgtgc aagtagaaag
     2341 ccagcctgtg gaaagcactg gcgctttcca aaggagacaa gggcccaggg actctacagc
     2401 caggagaacc cttgagctag ctccggagcg gccgcgtcga taatcaacct ctggattaca
     2461 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2521 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2581 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2641 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     2701 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     2761 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     2821 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga
     2881 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     2941 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3001 gtcggatctc cctttgggcc gcctccccgc atcgcggccg cggggatcca gacatgataa
     3061 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     3121 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     3181 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     3241 cggatcctct agagtcgacc tgcaggcatg caagctctag agcatggcta cgtagataag
     3301 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     3361 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3421 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc
     3481 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg attccgttgc
     3541 aatggctggc ggtaatattg ttctggatat taccagcaag gccgatagtt tgagttcttc
     3601 tactcaggca agtgatgtta ttactaatca aagaagtatt gcgacaacgg ttaatttgcg
     3661 tgatggacag actcttttac tcggtggcct cactgattat aaaaacactt ctcaggattc
     3721 tggcgtaccg ttcctgtcta aaatcccttt aatcggcctc ctgtttagct cccgctctga
     3781 ttctaacgag gaaagcacgt tatacgtgct cgtcaaagca accatagtac gcgccctgta
     3841 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3901 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3961 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4021 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4081 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4141 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4201 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4261 acaaaatatt aacgcttaca atttaaatat ttgcttatac aatcttcctg tttttggggc
     4321 ttttctgatt atcaaccggg gtacatatga ttgacatgct agttttacga ttaccgttca
     4381 tcgattctct tgtttgctcc agactctcag gcaatgacct gatagccttt gtagagacct
     4441 ctcaaaaata gctaccctct ccggcatgaa tttatcagct agaacggttg aatatcatat
     4501 tgatggtgat ttgactgtct ccggcctttc tcacccgttt gaatctttac ctacacatta
     4561 ctcaggcatt gcatttaaaa tatatgaggg ttctaaaaat ttttatcctt gcgttgaaat
     4621 aaaggcttct cccgcaaaag tattacaggg tcataatgtt tttggtacaa ccgatttagc
     4681 tttatgctct gaggctttat tgcttaattt tgctaattct ttgccttgcc tgtatgattt
     4741 attggatgtt ggaatcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     4801 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4861 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4921 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4981 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5041 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5101 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5161 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5221 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5281 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5341 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5401 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5461 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5521 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5581 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5641 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5701 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5761 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5821 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5881 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5941 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6001 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6061 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6121 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6181 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6241 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6301 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6361 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6421 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6481 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6541 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6601 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6661 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6721 gcctggtatc tttatagt
//
