LOCUS Exported 4904 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS EGFP-GalT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4904) AUTHORS Cole NB, Smith CL, Sciaky N, Terasaki M, Edidin M, Lippincott-Schwartz J TITLE Diffusional mobility of Golgi proteins in membranes of living cells. JOURNAL Science. 1996 Aug 9. 273(5276):797-801. PUBMED 8670420 REFERENCE 2 (bases 1 to 4904) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4904 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 84..387 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 388..591 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 542..562 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 867..876 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 873..1592 /codon_start=1 /product="enhanced GFP with monomerizing A206K mutation (Zacharias et al., 2002)" /label=mEGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(918..939) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1179..1198) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1526..1547 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1715..1836 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1752..1771) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1806..1825 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(1843..2298) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1980..2001) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2192..2211 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2325..2429 /gene="bla" /label=AmpR promoter promoter 2431..2788 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2454..2474) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 2639..2774 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2701..2720 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2823..3617 /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(2877..2896) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3487..3506 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3805..3824) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3849..3896 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4225..4813 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4714..4733 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt 61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg 601 aaccgtcaga tccgctagcg ctaccggact cagatctcga gctcgccacc atgaggcttc 661 gggagccgct cctgagcggc agcgccgcga tgccaggcgc gtccctacag cgggcctgcc 721 gcctgctcgt ggccgtctgc gctctgcacc ttggcgtcac cctcgtttac tacctggctg 781 gccgcgacct gagccgcctg ccccaactgg tcggagtctc cacaccgctg cagtcgacgg 841 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca 901 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg 961 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca 1021 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc 1081 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc 1141 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc 1201 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg 1261 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca 1321 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc 1381 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg 1441 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca 1501 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga 1561 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag 1621 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa 1681 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg 1741 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc 1801 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata 1861 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg 1921 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc 1981 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa 2041 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt 2101 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac 2161 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta 2221 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg 2281 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt 2341 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa 2401 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt 2461 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2521 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 2581 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 2641 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 2701 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg 2761 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc 2821 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 2881 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 2941 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3001 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3061 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3121 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3181 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3241 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3301 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg 3361 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3421 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 3481 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 3541 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 3601 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 3661 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 3721 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 3781 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg 3841 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa 3901 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg 3961 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc 4021 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat 4081 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt 4141 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 4201 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 4261 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 4321 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 4381 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 4441 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 4501 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 4561 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 4621 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 4681 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 4741 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 4801 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 4861 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtgg //