LOCUS Exported 4763 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION a mammalian expression backbone vector for humanized mNeonGreen. ACCESSION . VERSION . KEYWORDS pmNeonGreenHO-G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4763) AUTHORS Tanida-Miyake E, Koike M, Uchiyama Y, Tanida I TITLE Optimization of mNeonGreen for Homo sapiens increases its fluorescent intensity in mammalian cells. JOURNAL PLoS One. 2018 Jan 17;13(1):e0191108. doi: 10.1371/journal.pone.0191108. eCollection 2018. PUBMED 29342181 REFERENCE 2 (bases 1 to 4763) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4763 /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 537..546 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 1311..1367 /label=MCS /note="multiple cloning site" polyA_signal 1491..1612 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1528..1547) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1582..1601 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1619..2074) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1756..1777) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1968..1987 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2101..2205 /gene="bla" /label=AmpR promoter promoter 2207..2564 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2230..2250) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2415..2550 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2477..2496 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2599..3393 /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(2653..2672) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3263..3282 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3581..3600) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3625..3672 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4001..4589 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4490..4509 /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 gctagcgcca 541 ccatggtcag caaaggcgaa gaagacaaca tggcatcact gccagcgaca cacgaacttc 601 atattttcgg ctccatcaat ggtgtggact tcgatatggt tggacagggt actgggaacc 661 ctaacgatgg atatgaggag ttgaacttga agagtaccaa gggcgatctt cagttttcac 721 cttggattct cgtacctcat ataggttatg gctttcatca atatctgccc tatcctgacg 781 gtatgtcacc tttccaagca gctatggttg acgggagcgg ttatcaggtt caccggacaa 841 tgcagtttga agatggagcc tctttgaccg taaattatcg gtatacttat gagggcagcc 901 atattaaagg cgaggctcaa gttaagggga ctgggttccc cgcggatggt cccgtaatga 961 caaacagtct gacggctgca gactggtgcc gctctaagaa gacatatcca aatgacaaga 1021 cgattattag cactttcaag tggtcataca ctacaggaaa tggtaaacgc tataggagca 1081 ctgcccggac cacgtatact ttcgcgaagc cgatggcggc aaattatttg aaaaaccaac 1141 cgatgtatgt gttcagaaag acagaactca agcattctaa aactgaactg aacttcaaag 1201 agtggcagaa agcctttact gatgtaatgg gcatggacga actctataag tccggactca 1261 gatccggtgg aggcggttca ggcggaggtg gctctggcgg tggcggatcg agatctcgag 1321 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat 1381 aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc 1441 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta 1501 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 1561 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa 1621 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt 1681 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata 1741 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac 1801 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa 1861 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc 1921 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg 1981 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca 2041 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg 2101 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 2161 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga 2221 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca 2281 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct 2341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc 2401 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg 2461 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc 2521 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca 2581 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 2641 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 2701 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 2761 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc 2821 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 2881 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 2941 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 3001 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 3061 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 3121 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 3181 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 3241 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 3301 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 3361 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg 3421 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa 3481 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc 3541 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa 3601 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg 3661 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac 3721 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa 3781 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct 3841 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct 3901 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 3961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 4021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 4081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 4141 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 4201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 4261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 4321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 4381 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 4441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 4501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 4561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 4621 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 4681 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt 4741 catagcccat atatggagtt ccg //