LOCUS Exported 8910 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS hNICD3(3xFLAG)-pCDF1-MCS2-EF1-copGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8910) AUTHORS Zhao N, Liu H, Lilly B TITLE Reciprocal regulation of syndecan-2 and Notch signaling in vascular smooth muscle cells. JOURNAL J Biol Chem. 2012 May 11;287(20):16111-20. Epub 2012 Mar 21. PUBMED 22437834 REFERENCE 2 (bases 1 to 8910) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8910 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1217..1420 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1417..1441 /label=LNCX /note="Human CMV promoter, forward primer" CDS 1542..1607 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" promoter 3699..3910 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3923..4191 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" regulatory 4220..4229 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4250..4912 /codon_start=1 /product="green fluorescent protein 2 from Pontellina plumata, also known as ppluGFP2 (Shagin et al., 2004)" /label=CopGFP /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA" misc_feature 4991..5579 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5044..5064) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5462..5473) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5916..6037 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5953..5972) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6007..6026 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6077..6212 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6139..6158 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6245..6261) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6245..6261) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6258..6280) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6269..6285 /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(6293..6323) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6338..6359 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6476..6493) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6647..7235) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6727..6746) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7406..8266) /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 8029..8048 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8267..8371) /gene="bla" /label=AmpR promoter primer_bind 8439..8457 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8495..8517) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8617..8636 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 8781..74 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" ORIGIN 1 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct 61 atataagcag agcttgtgaa acttcgagga gtctctttgt tgaggacttt tgagttctcc 121 cttgaggctc ccacagatac aataaatatt tgagattgaa ccctgtcgag tatctgtgta 181 atctttttta cctgtgaggt ctcggaatcc gggccgagaa cttcgcagtt ggcgcccgaa 241 cagggacttg attgagagtg attgaggaag tgaagctaga gcaatagaaa gctgttaagc 301 agaactcctg ctgacctaaa tagggaagca gtagcagacg ctgctaacag tgagtatctc 361 tagtgaagca gactcgagct cataatcaag tcattgttta aaggcccaga taaattacat 421 ctggtgactc ttcgcggacc ttcaagccag gagattcgcc gagggacagt caacaaggta 481 ggagagattc tacagcaaca tggggaatgg acaggggcga gattggaaaa tggccattaa 541 gagatgtagt aatgttgctg taggagtagg ggggaagagt aaaaaatttg gagaagggaa 601 tttcagatgg gccattagaa tggctaatgt atctacagga cgagaacctg gtgatatacc 661 agagacttta gatcaactaa ggttggttat ttgcgattta caagaaagaa gagaaaaatt 721 tggatctagc aaagaaattg atatggcaat tcctgcattg aggagaaatg gtaggcaatg 781 tggcatgtct gaaaaagagg aggaatgatg aagtatctca gacttatttt ataagggaga 841 tactgtgctg agttcttccc tttgaggaag gtatgtcata tcctagacat agtctcaatt 901 ttaaaagaag aggtaggata ggagggatgg ccccttatga attattagca caacaagaat 961 ccttaagaat acaagattat ttttctgcaa taccacaaaa attgcaagca cagtggattt 1021 attataaaga tcaaaaagat aagaaatgga aaggaccaat gagagtagaa tactggggac 1081 agggatcagt attattaaag gatgaagaga agggatattt tcttataatc gatactagta 1141 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 1201 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 1261 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 1321 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 1381 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat 1441 cgcctggaga cgccatccac gctgttttga cctccataga agattctaga gcccgggcgc 1501 gccggatcca gatcttaatt aatttaaatg aattcaccat ggactacaaa gaccatgacg 1561 gtgattataa agatcatgat atcgattaca aggatgacga tgacaagctt gcatgcctgc 1621 aggtcgactc tagaatggtg gcccggcgca agcgcgagca cagcaccctc tggttccccg 1681 agggcttctc actgcacaag gacgtggcct ctggtcacaa gggccggcgg gaacccgtgg 1741 gccaggacgc gctgggcatg aagaacatgg ccaagggtga gagcctgatg ggggaggtgg 1801 ccacagactg gatggacaca gagtgcccag aggccaagcg gctaaaggta gaggagccag 1861 gcatgggggc tgaggaggct gtggattgcc gtcagtggac tcaacaccat ctggttgctg 1921 ctgacatccg cgtggcacca gccatggcac tgacaccacc acagggcgac gcagatgctg 1981 atggcatgga tgtcaatgtg cgtggcccag atggcttcac cccgctaatg ctggcttcct 2041 tctgtggggg ggctctggag ccaatgccaa ctgaagagga tgaggcagat gacacatcag 2101 ctagcatcat ctccgacctg atctgccagg gggctcagct tggggcacgg actgaccgta 2161 ctggcgagac tgctttgcac ctggctgccc gttatgcccg tgctgatgca gccaagcggc 2221 tgctggatgc tggggcagac accaatgccc aggaccactc aggccgcact cccctgcaca 2281 cagctgtcac agccgatgcc cagggtgtct tccagattct catccgaaac cgctctacag 2341 acttggatgc ccgcatggca gatggctcaa cggcactgat cctggcggcc cgcctggcag 2401 tagagggcat ggtggaagag ctcatcgcca gccatgctga tgtcaatgct gtggatgagc 2461 ttgggaaatc agccttacac tgggctgcgg ctgtgaacaa cgtggaagcc actttggccc 2521 tgctcaaaaa tggagccaat aaggacatgc aggatagcaa ggaggagacc cccctattcc 2581 tggccgcccg cgagggcagc tatgaggctg ccaagctgct gttggaccac tttgccaacc 2641 gtgagatcac cgaccacctg gacaggctgc cgcgggacgt agcccaggag agactgcacc 2701 aggacatcgt gcgcttgctg gatcaaccca gtgggccccg cagccccccc ggtccccacg 2761 gcctggggcc tctgctctgt cctccagggg ccttcctccc tggcctcaaa gcggcacagt 2821 cggggtccaa gaagagcagg aggccccccg ggaaggcggg gctggggccg caggggcccc 2881 gggggcgggg caagaagctg acgctggcct gcccgggccc cctggctgac agctcggtca 2941 cgctgtcgcc cgtggactcg ctggactccc cgcggccttt cggtgggccc cctgcttccc 3001 ctggtggctt cccccttgag gggccctatg cagctgccac tgccactgca gtgtctctgg 3061 cacagcttgg tggcccaggc cgggcgggtc tagggcgcca gccccctgga ggatgtgtac 3121 tcagcctggg cctgctgaac cctgtggctg tgcccctcga ttgggcccgg ctgcccccac 3181 ctgcccctcc aggcccctcg ttcctgctgc cactggcgcc gggaccccag ctgctcaacc 3241 cagggacccc cgtctccccg caggagcggc ccccgcctta cctggcagtc ccaggacatg 3301 gcgaggagta cccggcggct ggggcacaca gcagcccccc aaaggcccgc ttcctgcggg 3361 ttcccagtga gcacccttac ctgaccccat cccccgaatc ccctgagcac tgggccagcc 3421 cctcacctcc ctccctctca gactggtccg aatccacgcc tagcccagcc actgccactg 3481 gggccatggc caccaccact ggggcactgc ctgcccagcc acttcccttg tctgttccca 3541 gctcccttgc tcaggcccag acccagctgg ggccccagcc ggaagttacc cccaagaggc 3601 aagtgttggc ctgagacgct cgtcagttct tagatcttaa ttaatttaaa tgaattcgcg 3661 gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg gcagagcgca catcgcccac 3721 agtccccgag aagttggggg gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg 3781 cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg 3841 agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc 3901 cagaacacag ctgaagcttc gaggggctcg catctctcct tcacgcgccc gccgccctac 3961 ctgaggccgc catccacgcc ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc 4021 ctgaactgcg tccgccgtct aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc 4081 ggcgctccct tggagcctac ctagactcag ccggctctcc acgctttgcc tgaccctgct 4141 tgctcaactc tacgtctttg tttcgttttc tgttctgcgc cgttacagat ccaagctgtg 4201 accggcgcct acgctagacg ccaccatgga gagcgacgag agcggcctgc ccgccatgga 4261 gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg 4321 agagggcacc cccaagcagg gccgcatgac caacaagatg aagagcacca aaggcgccct 4381 gaccttcagc ccctacctgc tgagccacgt gatgggctac ggcttctacc acttcggcac 4441 ctaccccagc ggctacgaga accccttcct gcacgccatc aacaacggcg gctacaccaa 4501 cacccgcatc gagaagtacg aggacggcgg cgtgctgcac gtgagcttca gctaccgcta 4561 cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc accggcttcc ccgaggacag 4621 cgtgatcttc accgacaaga tcatccgcag caacgccacc gtggagcacc tgcaccccat 4681 gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta 4741 ctacagcttc gtggtggaca gccacatgca cttcaagagc gccatccacc ccagcatcct 4801 gcagaacggg ggccccatgt tcgccttccg ccgcgtggag gagctgcaca gcaacaccga 4861 gctgggcatc gtggagtacc agcacgcctt caagaccccc attgccttcg ccagatcccg 4921 cgctcagtcg tccaattctg ccgtggacgg caccgccgga cccggctcca ccggatctcg 4981 ctaagtcgac aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa 5041 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat 5101 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta 5161 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc 5221 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt 5281 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg 5341 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc 5401 ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc 5461 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct 5521 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc 5581 tggtaccgat gacagagtta gaagatcgct tcaggaagct atttggcacg acttctacaa 5641 cgggagacag cacagtagat tctgaagatg aacctcctaa aaaagaaaaa agggtggact 5701 gggatgagta ttggaaccct gaaatcgata gcttccagtg ctttgtgaaa cttcgaggag 5761 tctctttgtt gaggactttt gagttctccc ttgaggctcc cacagataca ataaatattt 5821 gagattgaac cctgtcgagt atctgtgtaa tcttttttac ctgtgaggtc tcggaatccg 5881 ggccgagaac ttcgcagcga gctcattgta ccgcgaactt gtttattgca gcttataatg 5941 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 6001 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc 6061 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 6121 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt 6181 ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacc aaattcgtaa 6241 tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac 6301 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 6361 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 6421 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 6481 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 6541 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 6601 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 6661 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 6721 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 6781 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 6841 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 6901 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 6961 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 7021 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 7081 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 7141 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 7201 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 7261 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 7321 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 7381 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 7441 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 7501 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 7561 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 7621 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 7681 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 7741 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 7801 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 7861 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 7921 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 7981 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 8041 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 8101 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 8161 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 8221 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 8281 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 8341 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 8401 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct 8461 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga 8521 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag 8581 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga 8641 gagtgcacca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 8701 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 8761 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 8821 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 8881 ggcaccaaaa tcaacgggac tttccaaaat //