LOCUS Exported 9936 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: Tight Junctions, Excitation: 604, Emission: 659. ACCESSION . VERSION . KEYWORDS mCardinal-ZO1-C-14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9936) AUTHORS Chu J, Haynes RD, Corbel SY, Li P, Gonzalez-Gonzalez E, Burg JS, Ataie NJ, Lam AJ, Cranfill PJ, Baird MA, Davidson MW, Ng HL, Garcia KC, Contag CH, Shen K, Blau HM, Lin MZ TITLE Non-invasive intravital imaging of cellular differentiation with a bright red-excitable fluorescent protein. JOURNAL Nat Methods. 2014 Mar 16. doi: 10.1038/nmeth.2888. PUBMED 24633408 REFERENCE 2 (bases 1 to 9936) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9936 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 547..556 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 553..1284 /codon_start=1 /product="far-red fluorescent protein derived from mNeptune, with further red-shifted spectra" /label=mCardinal /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV ARYCDLPSKLGHKLNGMDELYK" RBS 2985..2993 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" polyA_signal 6664..6785 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6701..6720) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6755..6774 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(6792..7247) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6929..6950) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7141..7160 /label=F1ori-R /note="F1 origin, reverse primer" promoter 7274..7378 /gene="bla" /label=AmpR promoter promoter 7380..7737 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(7403..7423) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 7588..7723 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7650..7669 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7772..8566 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7826..7845) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8436..8455 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(8754..8773) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 8798..8845 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 9174..9762 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9663..9682 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 541 ccggtcgcca ccatggtgag caagggcgag gagctgatca aggagaacat gcacatgaag 601 ctgtacatgg aaggcaccgt gaacaaccac cacttcaagt gcaccaccga aggggagggc 661 aagccctacg agggcaccca gacccagagg attaaggtgg tggagggagg ccccctgccg 721 ttcgcattcg acatcctggc cacctgcttt atgtacggga gcaagacctt catcaaccac 781 acccagggca tccccgattt ctttaagcag tccttccctg agggcttcac atgggagaga 841 gtcaccacat acgaagacgg gggcgtgctt accgttaccc aggacaccag cctccaggac 901 ggctgcttga tctacaacgt caagctcaga ggggtgaact tcccatccaa cggccctgtg 961 atgcagaaga aaacactcgg ctgggaggcc accaccgaga ccctgtaccc cgctgacggc 1021 ggcctggaag gcagatgcga catggccctg aagctcgtgg gcgggggcca cctgcactgc 1081 aacctgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc 1141 tactttgtgg accgcagact ggaaagaatc aaggaggccg acaatgagac ctacgtcgag 1201 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt 1261 aatggcatgg acgagctgta caagtccgga agtgctggga gcgcaagcgg aagcggaggc 1321 tccggtgagg aaacagctat atgggaacaa catacagtga cgcttcacag ggctcctgga 1381 tttggatttg gaattgcaat atctggtgga cgagataatc ctcattttca gagtggggaa 1441 acgtcaatag tgatttcaga tgtgctgaaa ggaggaccag ctgaaggaca gctacaggaa 1501 aatgaccgag ttgcaatggt taacggagtt tcaatggata atgttgaaca tgcttttgct 1561 gttcagcaac taaggaaaag tgggaaaaat gcaaaaatta caattagaag gaagaagaaa 1621 gttcaaatac cagtaagtcg tcctgatcct gaaccagtat ctgataatga agaagatagt 1681 tatgatgagg aaatacatga tccaagaagt ggccggagtg gtgtggttaa cagaaggagt 1741 gagaagattt ggccgaggga tagaagtgca agtagagaga ggagcttgtc cccgcggtca 1801 gacaggcggt cagtggcttc cagccagcct gctaaaccta ctaaagtcac actggtgaaa 1861 tcccggaaaa atgaagaata tggtcttcga ttggcaagcc atatatttgt taaggaaatt 1921 tcacaagata gtttggcagc aagagatggc aatattcaag aaggtgatgt tgtattgaag 1981 ataaatggta ctgtgacaga aaatatgtca ttgacagatg caaagacatt gatagaaagg 2041 tctaaaggca aattaaaaat ggtagttcaa agagatgaac gggctacgct attgaatgtc 2101 cctgatcttt ctgacagcat ccactctgct aatgcctctg agagagacga catttcagaa 2161 attcagtcac tggcatcaga tcattctggt cgatcacacg ataggcctcc ccgccgcagc 2221 cggtcacgat ctcctgacca gcggtcagag ccttctgatc attccaggca ctcgccgcag 2281 cagccaagca atggcagtct gcggagtaga gatgaagaga gaatttctaa acctggggct 2341 gtctcaactc ctgtaaagca tgctgatgat cacacaccta aaacagtgga agaagttaca 2401 gttgaaagaa atgagaaaca aacaccttct cttccagaac caaagcctgt gtatgcccaa 2461 gttggcaacc agatgtggat ttacctgtca gtccatctga tggtgtccta cctaattcaa 2521 ctcatgaaga tgggatttct tcggcccagc atgaaattgg taaaattcag aaaaggagat 2581 agtgtgggtt tgcggctggc tggtggaaat gatgttggaa tatttgtagc tggcgttcta 2641 gaagatagcc ctgcagccaa ggaaggctta gaggaaggtg atcaaattct cagggtaaac 2701 aacgtagatt ttacaaatat cataagagaa gaagccgtcc ttttcctgct tgacctccct 2761 aaaggagaag aagtgaccat attggctcag aagaagaagg atgtttatcg tcgcattgta 2821 gaatcagatg taggagattc tttctatatt agaacccatt ttgaatatga aaaggaatct 2881 ccctatggac ttagttttaa caaaggagag gtgttccgtg ctgtggatac cttgtacaat 2941 ggaaaactgg gctcttggct tgctattcga attggtaaaa atcataagga ggtagaacga 3001 ggcatcatcc ctaataagaa cagagctgag cagctcgcca gtgtacagta tacacttcca 3061 aaaacagcag gcggagaccg tgctgacttc tggagattca gaggtcttcg cagctccaag 3121 agaaatcttc gaaaaagcag agaggatttg tccgctcagc ctgttcaaac aaagtttcca 3181 gcttatgaaa gagtggttct tcgagaagct ggatttctga ggcctgtaac catttttgga 3241 ccaatagctg atgttgccag agaaaagctg gcaagagaag aaccagatat ttatcaaatt 3301 gcaaagagtg aaccacgaga cgctggaact gaccaacgta gctctggcta tattcgcctg 3361 catacaataa agcaaatcat agatcaagac aaacatgctt tattagatgt aacaccaaat 3421 gcagttgatc gtcttaacta tgcccagtgg tatccaattg ttgtatttct taaccctgat 3481 tctaagcaag gagtaaaaac aatgagaatg aggttatgtc cagaatctcg gaaaagtgcc 3541 aggaagttat acgagcgatc tcataaactt gctaaaaata atcaccatct ttttacaact 3601 acaattaact taaattcaat gaatgatggt tggtatggtg cgctgaaaga agcagttcaa 3661 caacagcaaa accagctggt atgggtttcc gagggaaagg cggatggtgc tacaagtgat 3721 gaccttgatt tgcatgatga tcgtctgtcc tacctgtcag ctccaggtag tgaatactca 3781 atgtatagca cggacagtag acacacttct gactatgaag acacagacac agaaggcggg 3841 gcctacactg atcaagaact agatgaaact cttaatgatg aggttgggac tccaccggag 3901 tctgccatta cacggtcctc tgagcctgta agagaggact cctctggaat gcatcatgaa 3961 aaccaaacat atcctcctta ctcaccacaa gcgcagccac aaccaattca tagaatagac 4021 tcccctggat ttaagccagc ctctcaacag aaagcagaag cttcatctcc agtcccttac 4081 ctttcgcctg aaacaaaccc agcatcatca acctctgctg ttaatcataa tgtaaattta 4141 actaatgtca gactggagga gcccacccca gctccttcca cctcttactc accacaagct 4201 gattctttaa gaacaccaag tactgaggca gctcacataa tgctaagaga tcaagaacca 4261 tcattgtcgt cgcatgtaga tccaacaaag gtgtatagaa aggacccata tcccgaggaa 4321 atgatgaggc agaaccatgt tttgaaacag ccagccgtta gtcacccagg gcacaggcca 4381 gacaaagagc ctaatctgac ctatgaaccc caactcccat acgtagagaa acaagccagc 4441 agagacctgg agcagcccac atacagatac gagtcctcaa gctatacgga ccagttttct 4501 cgaaactatg aacatcgtct gcgatacgaa gatcgcgtcc ccatgtatga agaacagtgg 4561 tcatattatg atgacaaaca gccctaccca tctcggccac cttttgataa tcagcactct 4621 caagaccttg actccagaca gcatcccgaa gagtcctcag aacgagggta ctttccacgt 4681 tttgaagagc cagcccctct gtcttacgac agcagaccac gttacgaaca ggcacctaga 4741 gcatccgccc tgcggcacga agagcagcca gctcctgggt atgacacaca tggtagactc 4801 agaccggaag cccagcccca cccttcagca gggcccaagc ctgcagagtc caagcagtat 4861 tttgagcaat attcacgcag ttacgagcaa gtaccacccc aaggatttac ctctagagca 4921 ggtcattttg agcctctcca tggtgctgca gctgtccctc cgctgatacc ttcatctcag 4981 cataagccag aagctctgcc ttcaaacacc aaaccactgc ctccaccccc aactcaaacc 5041 gaagaagagg aagatccagc aatgaagcca cagtctgtac tcaccagagt taagatgttt 5101 gaaaacaaaa ggtctgcatc cttagagacc aagaaggatg taaatgacac tggcagtttt 5161 aagcctccag aagtagcatc taaaccttca ggtgctccca tcattggtcc caaacccact 5221 tctcagaatc aattcagtga acatgacaaa actctgtaca ggattccaga acctcaaaaa 5281 cctcaactga agccacctga agatattgtt cggtccaatc attatgaccc tgaagaagat 5341 gaagaatatt atcgaaaaca gctgtcatac tttgaccgaa gaagttttga gaataagcct 5401 cctgcacaca ttgctgccag ccatctctcc gagcctgcaa agccagctca ttctcagaat 5461 caatcaaatt tttctagtta ttcttcaaag ggaaagcctc ctgaagctga tggtgtggat 5521 agatcatttg gcgagaaacg ctatgaaccc atccaggcca ctccccctcc tcctccattg 5581 ccctcgcagt atgcccagcc atctcagcct gtcaccagcg cgtctctcca catacattct 5641 aagggagcac atggtgaagg taattcagtg tcattggatt ttcagaattc cttagtgtcc 5701 aaaccagacc cacctccatc tcagaataag ccagcaactt tcagaccacc aaaccgagaa 5761 gatactgctc aggcagcttt ctatccccag aaaagttttc cagataaagc cccagttaat 5821 ggaactgaac agactcagaa aacagtcact ccagcataca atcgattcac accaaaacca 5881 tatacaagtt ctgcccgacc atttgaacgc aagtttgaaa gtcctaaatt caatcacaat 5941 cttctgccaa gtgaaactgc acataaacct gacttgtctt caaaaactcc cacttctcca 6001 aaaactcttg tgaaatcgca cagtttggca cagcctcctg agtttgacag tggagttgaa 6061 actttctcta tccatgcaga gaagcctaaa tatcaaataa ataatatcag cacagtgcct 6121 aaagctattc ctgtgagtcc ttcagctgtg gaagaggatg aagatgaaga tggtcatact 6181 gtggtggcca cagcccgagg catatttaac agcaatgggg gcgtgctgag ttccatagaa 6241 actggtgtta gtataattat ccctcaagga gccattcccg aaggagttga gcaggaaatc 6301 tatttcaagg tctgccggga caacagcatc cttccacctt tagataaaga gaaaggtgaa 6361 acactgctga gtcctttggt gatgtgtggt ccccatggcc tcaagttcct gaagcctgtg 6421 gagctgcgct taccacactg tgatcctaaa acctggcaaa acaagtgtct tcccggagat 6481 ccaaattatc tcgttggagc aaactgtgtt tctgtcctta ttgaccactt ttaaggatcc 6541 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct 6601 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt 6661 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc 6721 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta 6781 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 6841 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 6901 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 6961 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 7021 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 7081 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 7141 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 7201 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt 7261 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 7321 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc 7381 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc 7441 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga 7501 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 7561 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat 7621 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc 7681 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat 7741 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 7801 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 7861 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 7921 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct 7981 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 8041 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 8101 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 8161 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 8221 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 8281 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 8341 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 8401 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 8461 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 8521 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 8581 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc 8641 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc 8701 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca 8761 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa 8821 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct 8881 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca 8941 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc 9001 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa 9061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 9121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 9181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 9241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 9301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 9361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 9421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 9481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 9541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 9601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 9661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 9721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 9781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 9841 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac 9901 ggggtcatta gttcatagcc catatatgga gttccg //