LOCUS Exported 6738 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Plasmid encodes full-length (unprocessed) murine VEGF-D to be used for stimulation of lymphangiogenesis in mice in vivo.. ACCESSION . VERSION . KEYWORDS psubCMV-mVEGF-D-WPRE 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6738 /organism="synthetic DNA construct" /mol_type="other DNA" 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 4909..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" primer_bind 6682..6699 /label=L4440 /note="L4440 vector, 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 //