LOCUS Exported 5917 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses a TFR and ST6GAL1 swapped chimera with a truncated cytosolic tail from TFR, and TMD and luminal domain from ST6GAL1 in mammalian cells. ACCESSION . VERSION . KEYWORDS F9SS-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5917) TITLE Quantitative study of the Golgi retention of glycosyltranferases JOURNAL Unpublished REFERENCE 2 (bases 1 to 5917) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5917 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 113..416 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 417..620 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 571..591 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 671..680 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 1910..1919 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1916..2635 /codon_start=1 /product="monomeric GFP variant optimized for use in oxidizing environments such as the secretory pathway (Costantini et al., 2015)" /label=moxGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL KFISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(1961..1982) /label=EGFP-N /note="EGFP, reverse primer" polyA_signal 2757..2878 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2794..2813) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2848..2867 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2885..3340) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3022..3043) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3234..3253 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3367..3471 /gene="bla" /label=AmpR promoter promoter 3473..3830 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3496..3516) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3681..3816 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3743..3762 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3865..4659 /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(3919..3938) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4529..4548 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4847..4866) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4891..4938 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5267..5855 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5756..5775 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt 61 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat 121 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa 181 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg 241 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc 301 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct 361 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga 421 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa 481 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc 541 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg 601 aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggactc 661 agatctcgag gccaccatgg ccaatgtcac aaaaccaaaa aggttcagct gctgcgtcct 721 ggtctttctt ctgtttgcag tcatctgtgt gtggaaggaa aagaagaaag ggagttacta 781 tgattccttt aaattgcaaa ccaaggaatt ccaggtgtta aagagtctgg ggaaattggc 841 catggggtct gattcccagt ctgtatcctc aagcagcacc caggaccccc acaggggccg 901 ccagaccctc ggcagtctca gaggcctagc caaggccaaa ccagaggcct ccttccaggt 961 gtggaacaag gacagctctt ccaaaaacct tatccctagg ctgcaaaaga tctggaagaa 1021 ttacctaagc atgaacaagt acaaagtgtc ctacaagggg ccaggaccag gcatcaagtt 1081 cagtgcagag gccctgcgct gccacctccg ggaccatgtg aatgtatcca tggtagaggt 1141 cacagatttt cccttcaata cctctgaatg ggagggttat ctgcccaagg agagcattag 1201 gaccaaggct gggccttggg gcaggtgtgc tgttgtgtcg tcagcgggat ctctgaagtc 1261 ctcccaacta ggcagagaaa tcgatgatca tgacgcagtc ctgaggttta atggggcacc 1321 cacagccaac ttccaacaag atgtgggcac aaaaactacc attcgcctga tgaactctca 1381 gttggttacc acagagaagc gcttcctcaa agacagtttg tacaatgaag gaatcctaat 1441 tgtatgggac ccatctgtat accactcaga tatcccaaag tggtaccaga atccggatta 1501 taatttcttt aacaactaca agacttatcg taagctgcac cccaatcagc ccttttacat 1561 cctcaagccc cagatgcctt gggagctatg ggacattctt caagaaatct ccccagaaga 1621 gattcagcca aaccccccat cctctgggat gcttggtatc atcatcatga tgacgctgtg 1681 tgaccaggtg gatatttatg agttcctccc atccaagcgc aagactgacg tgtgctacta 1741 ctaccagaag ttcttcgata gtgcctgcac gatgggtgcc taccacccgc tgctctatga 1801 gaagaatttg gtgaagcatc tcaaccaggg cacagatgag gacatctacc tgcttggaaa 1861 agccacactg cctggcttcc ggaccattca ctgccgggat ccaccggtcg ccaccatggt 1921 gtccaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga 1981 cgtaaacggc cacaagttct ccgtgcgggg cgagggcgag ggcgatgcca ccaacggcaa 2041 gctgaccctg aagttcatca gcaccaccgg caagctgccc gtgccctggc ccaccctcgt 2101 gaccaccctg acctacggcg tgcagagctt ctcccgctac cccgaccaca tgaagcgcca 2161 cgacttcttc aagagcgcca tgcccgaagg ctacgtccag gagcgcacca tctccttcaa 2221 ggacgacggc acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa 2281 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct 2341 ggagtacaac ttcaactccc acaacgtcta tatcaccgcc gacaagcaga agaacggcat 2401 caaggccaac ttcaagatcc gccacaacgt ggaggacggc tccgtgcagc tcgccgacca 2461 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct 2521 gtccacccag tccaagctgt ccaaagaccc caacgagaag cgcgatcaca tggtccttct 2581 ggaattcgtg accgccgccg ggatcactca cggcatggac gagctgtaca agtaagcggc 2641 cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa 2701 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact 2761 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 2821 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag 2881 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc 2941 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc 3001 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac 3061 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca 3121 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg 3181 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag 3241 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc 3301 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga 3361 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 3421 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg 3481 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3541 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3601 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3661 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3721 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc 3781 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa 3841 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg 3901 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct 3961 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac 4021 ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg 4081 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg 4141 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa 4201 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca 4261 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt 4321 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc 4381 aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc 4441 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg 4501 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt 4561 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag 4621 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa 4681 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct 4741 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg 4801 gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg 4861 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg 4921 gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata 4981 ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc 5041 ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca 5101 tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 5161 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 5221 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 5281 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 5341 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 5401 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 5461 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 5521 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 5581 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 5641 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 5701 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 5761 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 5821 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 5881 ggttcctggc cttttgctgg ccttttgctc acatgtt //