LOCUS Exported 8299 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expression construct. ACCESSION . VERSION . KEYWORDS pCS-flt4um159-HA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8299) AUTHORS Shin M, Male I, Beane TJ, Villefranc JA, Kok FO, Zhu LJ, Lawson ND TITLE Vegfc acts through ERK to induce sprouting and differentiation of trunk lymphatic progenitors. JOURNAL Development. 2016 Oct 15;143(20):3785-3795. Epub 2016 Sep 12. PUBMED 27621059 REFERENCE 2 (bases 1 to 8299) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8299 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 35..53 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 35..52 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 108..132 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(4231..4255) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 4257..4283 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind complement(4288..4308) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter complement(4326..4343) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 4348..4482 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4372..4391) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 4426..4445 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(4595..4615) /label=T3 /note="T3 promoter, forward primer" promoter complement(4595..4613) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(4634..4650) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4634..4650) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4647..4669) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4658..4674 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4682..4712) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4727..4748 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4865..4882) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5036..5624) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5116..5135) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5795..6655) /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 6418..6437 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6656..6760) /gene="bla" /label=AmpR promoter rep_origin complement(6786..7241) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6923..6944) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7135..7154 /label=F1ori-R /note="F1 origin, reverse primer" promoter 7315..8299 /label=CMV IE94 promoter /note="enhancer/promoter region of simian cytomegalovirus major immediate early transcription unit IE94" ORIGIN 1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag 61 ctacttgttc tttttgcagg atcccatcga ttcgaattca aggatcaaca agtttgtaca 121 aaaaagcagg ctccgaattc gcccttgcca ccatgaagag agattttacg tttttctgtc 181 ggatttggat tgggattccc ttcttctcag gtctggtgaa tgggttttct atgagtccac 241 ccacccttga caacaccaaa gaccagcttg tgattaacgc taacgacaca ctgaacatta 301 catgcagggg tcagcgaatt ttggactgga gctggcctga agagtctttg agtaaagtgg 361 agtttactga tcgccagggt cagcagtcac ccactgacac cccagggtat agagaaatca 421 ggctgaagga gtgtcaaggg gtggctggaa aaccctactg caagatctta atacttacca 481 atgcccaggc caatgactcg ggttattacc gctgcttcta caaggacatt aaagctgtta 541 ttgatggcac cacggcagcc agcatttttg tgtttgttcg agacccagag catccattca 601 tcaaaagggg agacaacgac atggagacca tcttcataac agactctgaa acacacattg 661 aagtaccatg tctggtttca gatcccgacc taaaagtcac tctcttttcg ttagtgccgt 721 atccggagcc tgtggatggc agtgtggtca cctggaataa taaaaagggt tggtcgattc 781 ccaggcatat cattcagaac acttccacct ttattggctt ctactgctcc atctcagtcc 841 aaaacagcca gcacacatca tcaatctatg ttgtccaggt cattgggctg aagttttatg 901 agttcaagct gtttcctgaa gactcgcctg tggagttgat gcagggggag agtttggtgc 961 tcaactgcac tgcactggtc gacttcaaca ctggtgtgga tttccagtgg gactatccag 1021 gcaaaaagga gaaccggttg gcgagtttac agcctctgcg taacgtcctg gatgaagcca 1081 cggaaatctc cagtatcctc tcaatcagaa acattcactt ggatgatagt ggctactaca 1141 cctgctgggc taatactctg gaaatgaaac gagagctcac tactgtagtc atagttcatg 1201 aaaaaccttt tatcagcttg gactacagga atggatcagt tatagaggca aaagagggac 1261 aaaaatctgt ccggctatcc gtaaaagtgt cagcataccc ttcaccagaa atacaatggt 1321 ataagaatgg aaaactgatc agcagcaaga attcaagcag gtttaaggtc cagcagcact 1381 ctctacagat cagggatgtg tgtaagcagg atgctggaga atacatgctg gtcctgaaaa 1441 atagccctgc agccctggag aaaagactca actttactct catagtcaac gttccacctc 1501 agattcatga gaaggaggca gcaccaccta caaacctcta tgggaagggc actcgacaga 1561 tcctcacttg caccgcagac gggagccctc cagcctccat cagctggcag tggaggccct 1621 ggagcccctg cgatctggag cgtacccgca gagccctgag acggaggggt ggacgagacc 1681 agagtccttt ctgtcacaac tggatggatt tggaccctga acatgctgta aaccctattg 1741 agagtattga cacattgact cagatggtgg acggcaaaga aaagactgtc ggccgtgtgg 1801 ttattcagaa tgccagtgtg ccagctatgt tggctgagaa caaagtggga aaagatgaac 1861 gactgattta tttctacgta accactatcc ctgaaggatt cgatatagag atggagccct 1921 cagaggatcc gcttgagcag gacctggtac agctgaagtg taatgcagat aatttcacct 1981 atgagaacct gcggtggtac cgcctggacc cgcagactgt tcctccagag ctggactgca 2041 agagtctgca ccagtatgcc acatttttgg agggacaact atcttttcag accaccagca 2101 acaactgggt cctgcagctt aacattacca acattcagct acaggatgag gggaactatg 2161 tgtgtgaggt gcagaaccga cgaaccgggg tgaaacattg tcaccgcaaa tacatcccag 2221 tcaaagcaat ggaagcacct cgataccgtc acaacccaac taatcatacg gtaaatgtga 2281 gcgagtcgct gcagatgaac tgtgatgtgg aaggcacacc ctttcctcag ctgtcctggt 2341 ttaaagataa ccaacccctc catcagatat cagggatcct ccttcaggac tccaatcgga 2401 ctctcagcat tcagcgagtg cgagaggaag acgccggtct gtacacctgc tctgcctgca 2461 accagaaggg ctgcgttcag tcctccgcca cggtgtcagt gattggctct gatgacaaaa 2521 ccaatgtaga aatagtgatc ctcatcggca caggagtcat cgccatcttt ttctgggtcc 2581 ttcttcttgt catcttctgc aatgtcaagc gagtgaaccc agctgatatc aagacaggct 2641 atctgtccat catcatggac ccgggggaag tgccgctgga ggagcagtgt gaatacctgc 2701 cgtacgactc cagccagtgg gagatttcca gagacagact gcgcctcggt aaggtgctcg 2761 gccatggagc tttcgggaag gtgattgaag cctcaatctt tggtcatgac aagaaatcct 2821 ctgcaaacac agtggctgtc aagatgttga aagagggagc aacagccagc gagcacaaag 2881 ccctgatgtc agagctgaag atcctcattc atattggaaa ccatctcaat gttgtcaacc 2941 ttcttggagc ttgtaccaaa cctaacggtc ctctcatggt tatagtggag tattgtaaat 3001 atggaaatct gtccaacttc ctgcgtgcca agagggagtt ttttttacca tacagggatc 3061 gttctcctaa aacccaaagt caagtgcgac ggatgataga agcaggtcag gcgagccaga 3121 gtgagcatca gccgtccact tcctccacca accctccacg tgttacagtg gatgatctct 3181 ggaaaactcc actcacaata gaggatctga tatgctacag ttttcaagtt gcacgaggaa 3241 tggagtttct ggcatctcgt aagtgtattc atcgagatct ggcagcccga aacatcctcc 3301 tgtccgagaa caatgtggtg aagatctgtg actttggttt agcacgggac atctacaaag 3361 accctgacta tgtgcgcaaa ggcaatgcca ggctgccgct gaagtggatg gctccagaga 3421 gcatctttga taaggtttac accagtcaga gtgacgtctg gtcttttgga gttctgctct 3481 gggagatttt ctcactagga gcgtctccat accccggcat ccagattgat gaagatttct 3541 gcaaacgact caaagacggt accaggatga gagcaccaga caatgcgtcc cctgaaatat 3601 atggcatcat gttggcctgc tggcagggcg agcccaggga gagacccaca tttccagctc 3661 tggtggagat acttggagat ctactgcagg aaaacagtct accagaaatc ccgttcaatg 3721 tgtctcagag ctcagaggac gatggcttct cacaagcttc ttccagacca ccatcacaag 3781 aggagatcag attggcctgc aacacactgc caacacgata ttacaactgc gtgccatttg 3841 ctggctgtgt gatggtcggg ccctccagca catgtcattc tcgagtgaag acctttgagg 3901 aacttcctat ggaaatgaca tcacacaaga cacagcatga cagtcagaca gacagtggga 3961 tggttttggc atcagatgaa ctggagaggt ttgaacacaa gcacagagga gccatgttga 4021 cacatgcgac gacgggccaa agcactgagc gcctcatcag ttgtccatcg gtgagcagca 4081 gcggcagcgg tggtggtttg ctccgtccag tgttcttcac tcaactctca ggccagacct 4141 tctacaacaa cgagtacggt catctgtcag aggagggcgt cagcgactac ttctcctcct 4201 cagaccaggc cgtttacaag ggcgaattcg acccagcttt cttgtacaaa gtggactatc 4261 cgtacgacgt accagactac gcataaccaa ctttattata catagttgca ctcgagcctc 4321 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt 4381 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct 4441 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt 4501 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg 4561 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct 4621 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4681 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4741 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4801 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4861 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4921 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4981 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 5041 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 5101 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 5161 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 5221 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 5281 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 5341 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 5401 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 5461 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 5521 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 5581 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 5641 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5701 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5761 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5821 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5881 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5941 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 6001 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 6061 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 6121 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 6181 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 6241 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 6301 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 6361 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 6421 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 6481 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 6541 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 6601 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6661 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6721 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6781 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 6841 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 6901 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 6961 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 7021 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 7081 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 7141 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 7201 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct 7261 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccat 7321 agccaattca atatggcgta tatggactca tgccaattca atatggtgga tctggacctg 7381 tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga tctggacccc 7441 agccaattca atatggcgga cttggcacca tgccaattca atatggcgga cttggcactg 7501 tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg tcttggccct 7561 gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat attggctata 7621 tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg gctattggcc 7681 aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg gttttcctat 7741 tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg cttcctaata 7801 ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct ttcctattga 7861 cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt caattacggt 7921 aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc attgacgtca 7981 atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac gtcaatgacg 8041 gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc aagtacatta 8101 ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat ggcggtaaat 8161 ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg caatgacgca 8221 aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg tctatataag 8281 caatgctcgt ttagggaac //