LOCUS Exported 5758 bp ds-DNA circular SYN 16-OCT-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 5758) AUTHORS Joe TITLE Direct Submission JOURNAL Exported Monday, May 4, 2020 from SnapGene 5.1.0 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5758 /organism="recombinant plasmid" /mol_type="other DNA" primer_bind 1..28 /label=Fragment 1.FOR CDS 11..70 /codon_start=1 /label=Membrane Trafficking signal /note="Membrane trafficking signal" /translation="KSRITSEGEYIPLDQIDINV" primer_bind complement(50..70) /label=eViRhos_dC_r CDS 71..784 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(113..134) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(374..393) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 721..742 /label=EGFP-C /note="EGFP, forward primer" CDS 785..808 /codon_start=1 /label=ER release signal /translation="FCYENEV" primer_bind 806..838 /label=eViRhos_dC_f polyA_signal 919..1040 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(956..975) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1010..1029 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1047..1502) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1184..1205) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1396..1415 /label=F1ori-R /note="F1 origin, reverse primer" promoter 1529..1633 /gene="bla" /label=AmpR promoter promoter 1635..1992 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(1658..1678) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 1843..1978 /label=SV40 ori /note="SV40 origin of replication" primer_bind 1905..1924 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(1928..1939) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2027..2821 /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(2081..2100) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2691..2710 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3009..3028) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3053..3100 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 3429..4017 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3918..3937 /label=pBR322ori-F /note="pBR322 origin, forward primer" enhancer 4192..4495 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4496..4699 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4650..4670 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 4740..4796 /label=MCS /note="multiple cloning site" regulatory 4797..4806 /regulatory_class="other" /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4803..4934 /codon_start=1 /label=N-Terminus eKR2 /translation="MDYGGALSAVGLFQTSYTLENNGSVICIPNNGQCFCLAWLKSNG" primer_bind complement(4916..4944) /label=Fragment 1.REV primer_bind 4925..4957 /label=Fragment 2.FOR CDS join(4935..5758,1..10) /codon_start=1 /label=viRho2_dC /label=295 2620979_21821 /translation="MSFDQTICKHIQFIDLLLSIILFIFFYYQKIVKRRCKFEVLYVTG VGSLTDLVKLTITNELPASYVIENNGVLFNWSQFLGWMITCPVLLIHLSNLAGKDVFDV RRMMKILVIYQILMVSGATASMCNNDLKWLFFGVSLISLLKIYIYAHNIFQESREIMPK RANVVINWISFVFYVSWSGFGVSWFFSPNGISFFSDYTTKALFAFFDIMSKNVYSMLGW YLRWYIIRKFDNPEEFVEQYQEEDEKKINILLVENNAIYSHYFDNILIQHGCTVDV" primer_bind complement(join(5751..5758,1..20)) /label=Fragment 2.REV ORIGIN 1 cgtggacgtg aagagcagga tcaccagcga gggcgagtac atccccctgg accagatcga 61 catcaacgtg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga 121 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc 181 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg 241 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca 301 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 361 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 421 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 481 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 541 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 601 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 661 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca 721 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 781 caagttctgc tacgagaacg aggtgtgatc tagatcataa tcagccatac cacatttgta 841 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 901 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 961 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 1021 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc 1081 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 1141 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 1201 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 1261 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 1321 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 1381 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 1441 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 1501 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 1561 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 1621 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 1681 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 1741 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 1801 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 1861 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 1921 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 1981 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 2041 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 2101 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 2161 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 2221 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 2281 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 2341 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 2401 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 2461 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 2521 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 2581 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 2641 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 2701 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 2761 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 2821 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 2881 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 2941 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 3001 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 3061 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 3121 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 3181 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 3241 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 3301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 3361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 3541 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 3781 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 4081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta 4141 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 4201 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 4261 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 4321 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 4381 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 4441 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 4501 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 4561 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 4621 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 4681 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca 4741 gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg ggatccgcca 4801 ccatggacta tgggggagca ctgtccgccg tcgggctgtt tcagacttcc tacaccctgg 4861 agaacaacgg gagcgtcatt tgcattccta acaatggcca gtgcttctgt ctggcctggc 4921 tgaagagcaa cgggatgtcc tttgaccaga ccatttgtaa gcatattcag tttattgacc 4981 tgctgctgtc cattatcctg ttcatcttct tctattatca gaagatcgtg aagaggagat 5041 gcaagttcga ggtgctgtac gtgaccggcg tgggctctct gacagacctg gtgaagctga 5101 ccatcacaaa tgagctgcca gcctcctatg tgatcgagaa caatggcgtg ctgttcaact 5161 ggtctcagtt tctgggctgg atgatcacct gccccgtgct gctgatccac ctgagcaatc 5221 tggccggcaa ggacgtgttc gatgtgcggc gcatgatgaa gatcctggtc atctaccaga 5281 tcctgatggt gtccggcgcc acagcctcta tgtgcaacaa tgatctgaag tggctgttct 5341 ttggcgtgtc tctgatcagc ctgctgaaga tctacatcta tgcccacaac atctttcagg 5401 agagccggga gatcatgcct aagcgcgcca acgtggtcat caactggatc tccttcgtgt 5461 tttacgtgag ctggtccggc ttcggcgtga gctggttctt ttccccaaat ggcatctcct 5521 tcttttctga ctacaccaca aaggccctgt ttgccttctt tgatatcatg agcaagaacg 5581 tgtattccat gctgggctgg tatctgaggt ggtatatcat cagaaagttc gacaatcccg 5641 aggagtttgt ggagcagtac caggaggagg atgagaagaa gatcaacatc ctgctggtgg 5701 agaacaatgc catctactcc cactatttcg acaatatcct gatccagcac ggctgtac //