LOCUS       sequence-231678-        7895 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid encodes Strep-tagged full-length (unprocessed) VEGF-D of 
            human origin 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 7895)
  TITLE     Alitalo expression plasmids
REFERENCE   2  (bases 1 to 7895)
  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: psubCAG-WPRE-hVEGF-D-Strept
FEATURES             Location/Qualifiers
     source          1..7895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   35..164
                     /label=AAV2 ITR
     enhancer        223..602
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        604..881
                     /label=chicken beta-actin promoter
     intron          882..1898
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1822..1844
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1906..1925
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1989..2085
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             join(3219..3224,3225..3248,3249..3284,3285..3308)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     misc_feature    3322..3910
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3375..3395)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3793..3804)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3925..4059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3949..3968)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4003..4022
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4209..4338
                     /label=AAV2 ITR
     rep_origin      4709..5222
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4796..4815)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5006..5027
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5704..5723)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5823..5845
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5883..5901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5969..6073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6074..6142,6143..6934)
                     /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(6292..6311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7105..7693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7594..7613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctctccccgc gcgttggccg attcattaat gcagctgcgc gctcgctcgc tcactgaggc
       61 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      121 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      181 cccgccatgc tacttatcta cgtagccatg ctctagaatt tcgacattga ttattgacta
      241 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      301 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      361 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      421 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      481 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      541 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      601 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
      661 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
      721 gggggcgcgc gccrggsggg gsggggsggg gsgrggggsg gggsggggcg aggcggagag
      781 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      841 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      901 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      961 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     1021 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     1081 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     1141 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     1201 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     1261 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     1321 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     1381 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     1441 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     1501 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     1561 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     1621 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     1681 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     1741 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     1801 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     1861 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     1921 ttgtgctgtc tcatcatttt ggcaaagaat tcctcgatcg aggaactgaa aaaccagaaa
     1981 gttaactggt aagtttagtc tttttgtctt ttatttcagg tcccggatcc ggtggtggtg
     2041 caaatcaaag aactgctcct cagtggatgt tgcctttact tctaggcctg tacggaagtg
     2101 ttacttctgc tctaaaagct gcggaattgt acccggcggc cgctagcgcc accatgtaca
     2161 gagagtgggt agtggtgaat gttttcatga tgttgtacgt ccagctggtg cagggctcca
     2221 gtaatgaaca tggaccagtg aagcgatcat ctcagtccac attggaacga tctgaacagc
     2281 agatcagggc tgcttctagt ttggaggaac tacttcgaat tactcactct gaggactgga
     2341 agctgtggag atgcaggctg aggctcaaaa gttttaccag tatggactct cgctcagcat
     2401 cccatcggtc cactaggttt gcggcaactt tctatgacat tgaaacacta aaagttatag
     2461 atgaagaatg gcaaagaact cagtgcagcc ctagagaaac gtgcgtggag gtggccagtg
     2521 agctggggaa gagtaccaac acattcttca agcccccttg tgtgaacgtg ttccgatgtg
     2581 gtggctgttg caatgaagag agccttatct gtatgaacac cagcacctcg tacatttcca
     2641 aacagctctt tgagatatca gtgcctttga catcagtacc tgaattagtg cctgttaaag
     2701 ttgccaatca tacaggttgt aagtgcttgc caacagcccc ccgccatcca tactcaatta
     2761 tcagaagatc catccagatc cctgaagaag atcgctgttc ccattccaag aaactctgtc
     2821 ctattgacat gctatgggat agcaacaaat gtaaatgtgt tttgcaggag gaaaatccac
     2881 ttgctggaac agaagaccac tctcatctcc aggaaccagc tctctgtggg ccacacatga
     2941 tgtttgacga agatcgttgc gagtgtgtct gtaaaacacc atgtcccaaa gatctaatcc
     3001 agcaccccaa aaactgcagt tgctttgagt gcaaagaaag tctggagacc tgctgccaga
     3061 agcacaagct atttcaccca gacacctgca gctgtgagga cagatgcccc tttcatacca
     3121 gaccatgtgc aagtggcaaa acagcatgtg caaagcattg ccgctttcca aaggagaaaa
     3181 gggctgccca ggggccccac agccgaaaga atcctgggag cgcttggagc cacccacagt
     3241 tcgaaaaagg tggaggttct ggcggtggat cgggaggttc agcgtggtct catcctcaat
     3301 ttgagaagtg aacgcgtcga taatcaacct ctggattaca aaatttgtga aagattgact
     3361 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3421 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3481 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3541 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3601 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3661 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     3721 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3781 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3841 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3901 gcctccccgc atcgcggccg cggggatcca gacatgataa gatacattga tgagtttgga
     3961 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     4021 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     4081 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt cggatcctct agagtcgacc
     4141 tgcaggcatg caagctctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     4201 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4261 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4321 gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     4381 acagttgcgc agcctgaatg gcgaatggcg attccgttgc aatggctggc ggtaatattg
     4441 ttctggatat taccagcaag gccgatagtt tgagttcttc tactcaggca agtgatgtta
     4501 ttactaatca aagaagtatt gcgacaacgg ttaatttgcg tgatggacag actcttttac
     4561 tcggtggcct cactgattat aaaaacactt ctcaggattc tggcgtaccg ttcctgtcta
     4621 aaatcccttt aatcggcctc ctgtttagct cccgctctga ttctaacgag gaaagcacgt
     4681 tatacgtgct cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4741 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4801 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4861 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4921 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4981 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5041 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5101 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     5161 atttaaatat ttgcttatac aatcttcctg tttttggggc ttttctgatt atcaaccggg
     5221 gtacatatga ttgacatgct agttttacga ttaccgttca tcgattctct tgtttgctcc
     5281 agactctcag gcaatgacct gatagccttt gtagagacct ctcaaaaata gctaccctct
     5341 ccggcatgaa tttatcagct agaacggttg aatatcatat tgatggtgat ttgactgtct
     5401 ccggcctttc tcacccgttt gaatctttac ctacacatta ctcaggcatt gcatttaaaa
     5461 tatatgaggg ttctaaaaat ttttatcctt gcgttgaaat aaaggcttct cccgcaaaag
     5521 tattacaggg tcataatgtt tttggtacaa ccgatttagc tttatgctct gaggctttat
     5581 tgcttaattt tgctaattct ttgccttgcc tgtatgattt attggatgtt ggaatcgcct
     5641 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
     5701 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5761 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5821 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5881 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5941 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6061 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6121 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6181 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6241 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6301 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6361 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6421 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6481 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6541 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6601 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6661 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7861 cgaggaagcg gaagagcgcc caatacgcaa accgc
//
