LOCUS Exported 4731 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Localization: C1 Cloning Vector, Excitation: 384, Emission: 450. ACCESSION . VERSION . KEYWORDS mAzurite-C1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4731) AUTHORS Mena MA, Treynor TP, Mayo SL, Daugherty PS TITLE Blue fluorescent proteins with enhanced brightness and photostability from a structurally targeted library. JOURNAL Nat Biotechnol. 2006 Dec . 24(12):1569-71. PUBMED 17115054 REFERENCE 2 (bases 1 to 4731) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4731 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 133..436 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 437..640 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 591..611 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 679..688 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 685..1401 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSHGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNIYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFRTAAGITHGMDELYK" primer_bind complement(730..751) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(991..1010) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 1402..1467 /label=MCS /note="multiple cloning site" polyA_signal 1591..1712 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1628..1647) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1682..1701 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1719..2174) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1856..1877) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2068..2087 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2201..2305 /gene="bla" /label=AmpR promoter promoter 2307..2664 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2330..2350) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2515..2650 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2577..2596 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2699..3493 /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(2753..2772) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3363..3382 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3681..3700) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3725..3772 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4101..4689 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4590..4609 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 61 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata 121 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 181 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 241 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 301 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 361 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 421 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 481 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 541 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 601 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 661 ccgctagcgc taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg 721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 841 aagctgcccg tgccctggcc caccctcgtg accaccctga gccacggcgt gcagtgcttc 901 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 961 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 1021 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 1081 gaggacggca acatcctggg gcacaagctg gagtacaact tcaacagcca caacatctat 1141 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 1201 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 1261 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag caaagacccc 1321 aacgagaagc gcgatcacat ggtcctgctg gagttccgca ccgccgccgg gatcactcac 1381 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca 1441 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat 1501 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg 1561 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac 1621 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 1681 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt 1741 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat 1801 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt 1861 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt 1921 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag 1981 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg 2041 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc 2101 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc 2161 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt 2221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct 2281 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt 2341 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 2401 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2461 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg 2521 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt 2581 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt 2641 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat 2701 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 2761 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 2821 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 2881 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 2941 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 3001 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 3061 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 3121 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 3181 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 3241 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 3301 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 3361 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 3421 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 3481 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg 3541 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt 3601 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc 3661 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct 3721 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc 3781 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3841 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3901 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt 3961 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 4021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 4081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 4141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 4201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 4261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 4321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 4381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 4441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 4501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 4561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 4621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 4681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg c //