LOCUS Exported 5313 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION mammalian expression of Sar1b tagged with YFP. ACCESSION . VERSION . KEYWORDS Sar1b-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5313) TITLE ER to Golgi protein delivery through an interwoven, tubular network extending from ER JOURNAL Unpublished REFERENCE 2 (bases 1 to 5313) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5313 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 60..363 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 364..567 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 518..538 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1258..1977 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1303..1324) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1564..1583) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1911..1932 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 2100..2221 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2137..2156) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2191..2210 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2228..2683) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2365..2386) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2577..2596 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2710..2814 /gene="bla" /label=AmpR promoter promoter 2816..3173 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2839..2859) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3024..3159 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3086..3105 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3208..4002 /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(3262..3281) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3872..3891 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4190..4209) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4234..4281 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4610..5198 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5099..5118 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 agttattaat agtaatcaat tacggggtca ttagttaata gcccatatat ggagttccgc 61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 361 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 421 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 481 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 541 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac 601 cggactcaga tctcgagctc aagcttcgaa ttcaccgcca ccatgtcctt catatttgac 661 tggatttaca gtggtttcag cagtgtgctg cagtttctag gactgtataa aaaaactggt 721 aaactggtat tccttggatt ggacaatgct gggaaaacca cgttgctaca catgctgaaa 781 gatgacagac ttggccagca cgtcccaaca ctacatccca cttcagaaga actcactatt 841 gctggcatga cttttacaac ttttgatctg ggtgggcaca tccaagcccg aagagtatgg 901 aaaaattacc ttcctgctat caatggcatt gtatttcttg tggattgtgc agaccatgaa 961 aggctgttag aatcaaaaga agaacttgat tcactaatga cagatgaaac cattgccaat 1021 gtgcctatat taattcttgg aaataagatt gacagacccg aagccatcag tgaagagagg 1081 ttacgagaga tgtttggctt atatgggcag acaacaggaa agggcagtgt gtcactgaag 1141 gagctgaatg ccagacctct ggaagtgttc atgtgcagtg tgctgaaaag gcaaggctat 1201 ggagaaggct tccgctggat ggcacagtac atcgatgggg atccaccggt cgcccatatg 1261 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 1321 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 1381 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 1441 gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca catgaagcag 1501 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 1561 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 1621 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 1681 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 1741 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 1801 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 1861 ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 1921 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc 1981 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa 2041 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta 2101 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 2161 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 2221 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc 2281 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag 2341 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg 2401 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca 2461 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa 2521 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg 2581 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta 2641 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg 2701 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 2761 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag 2821 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 2881 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 2941 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3001 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3061 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 3121 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat 3181 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 3241 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 3301 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 3361 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc 3421 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 3481 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 3541 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 3601 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 3661 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 3721 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 3781 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 3841 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 3901 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 3961 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 4021 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 4081 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 4141 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga 4201 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 4261 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg 4321 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc 4381 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg 4441 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta 4501 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 4561 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 4621 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4681 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 4741 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 4801 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 4861 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 4921 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 4981 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 5041 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 5101 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 5161 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 5221 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 5281 attctgtgga taaccgtatt accgccatgc att //