LOCUS Exported 6953 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian expression of rat GAT with N terminal CFP fusion. ACCESSION . VERSION . KEYWORDS pECFP-C1-rGAT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6953) AUTHORS Scholze P, Freissmuth M, Sitte HH TITLE Mutations within an intramembrane leucine heptad repeat disrupt oligomer formation of the rat GABA transporter 1. JOURNAL J Biol Chem. 2002 Nov 15;277(46):43682-90. Epub 2002 Sep 9. PUBMED 12223478 REFERENCE 2 (bases 1 to 6953) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6953 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 607..616 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 613..1329 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(658..679) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(919..938) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1266..1287 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3403..3537 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" polyA_signal 3741..3862 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(3869..4324) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4006..4027) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4218..4237 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4351..4455 /gene="bla" /label=AmpR promoter promoter 4457..4814 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4480..4500) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4665..4800 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4727..4746 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4849..5643 /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(4903..4922) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5513..5532 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5831..5850) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5875..5922 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6251..6839 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6740..6759 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg 661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc 721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg 781 ccctggccca ccctcgtgac caccctgacc tggggcgtgc agtgcttcag ccgctacccc 841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag 901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag 961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac 1021 atcctggggc acaagctgga gtacaactac atcagccaca acgtctatat caccgccgac 1081 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga ggacggcagc 1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg 1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc 1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag 1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgagtggact 1381 ggagctgaca tcgcacgcct gcccgccagg atccctgctg ccaagttcgt gctccgagac 1441 atggcgactg acaacagcaa ggtggctgat gggcagatct ctaccgaggt cagcgaggct 1501 cctgtggcca gtgacaagcc caaaaccctt gtagtcaagg tgcagaagaa ggctggagac 1561 ctccctgacc gggacacatg gaagggacgc ttcgacttcc tcatgtcctg tgtgggctac 1621 gccatcggcc tgggcaatgt atggaggttc ccttatctct gcggaaagaa cggtggtggg 1681 gccttcctaa ttccatattt cctgacgctc atctttgcgg gtgttcctct cttccttttg 1741 gagtgctccc taggccagta cacctccatt gggggcctgg gcgtatggaa gctggctccc 1801 atgttcaagg gtgtgggcct cgcggcagct gtgctgtcct tctggctgaa catttattac 1861 attgtcatca tctcctgggc catctactac ctgtacaact ccttcaccac gactctgccg 1921 tggaaacagt gtgacaaccc ctggaacact gaccgctgct tctccaacta tagcctggtc 1981 aacaccacca acatgaccag tgccgtggtg gaattctggg agcgcaacat gcaccaaatg 2041 acagatgggc tagacaagcc aggacagatc cgctggccgc tagccatcac actggccatt 2101 gcctgggtgc ttgtgtattt ctgcatctgg aagggtgttg gatggactgg aaaggtggtc 2161 tacttctcag ccacataccc ctacatcatg ctcatcatcc tgttcttccg tggagtgaca 2221 cttcctgggg ccaaggaggg gatcctcttc tacatcacgc ccaacttccg aaagctgtct 2281 gattctgagg tgtggcttga cgccgccacc cagatcttct tctcctacgg actgggcctg 2341 gggtccctga ttgctctggg aagctacaac tctttccaca acaatgtgta cagggactcc 2401 atcattgttt gctgcatcaa ctcctgcacc agcatgtttg ccggcttcgt catcttctcc 2461 atcgtgggct tcatggctca tgtcaccaag aggtccatag ccgatgtggc ggcctcaggc 2521 cctggactgg cattcttggc ataccccgag gctgtgacac agctacccat ctctcccctc 2581 tgggctatcc tcttcttctc catgctgctg atgctgggca ttgacagcca gttctgtacc 2641 gtggagggct tcatcactgc cctggtggat gagtacccca ggcttctccg caatcgccgt 2701 gaactcttca ttgctgccgt gtgcatcgtg tcctacctga ttggcctctc taacatcacc 2761 cagggtggca tttatgtctt caaactgttc gattattact ctgccagtgg catgagcctg 2821 ctgttcctgg ttttctttga gtgtgtctcc atttcctggt tttatggtgt caaccggttc 2881 tatgacaaca tccaggagat ggttggctcc aggccctgca tctggtggaa gctgtgctgg 2941 tcctttttca cacccatcat tgtggcgggc gtgtttctct tcagtgctgt gcagatgaca 3001 ccactcacca tgggaagcta cgttttcccc aagtggggcc agggcgtggg ctggctcatg 3061 gctctgtcct ccatggtgct catccccggg tacatggctt acatgttcct caccctaaag 3121 ggctctctga agcagcgtct ccaggtcatg attcagccca gtgaagatat cgtgcgccct 3181 gagaatggtc ctgagcagcc gcaggctggc agctcagcca gcaaggaggc ctacatctag 3241 gggtacagcc ccctcatcac ccctacactg gcaccctgga ctggctgtac ccacacccct 3301 tgaagactga agatactctc tgtctccaac tacctcaagg ggcgggtcca gacaccatca 3361 ccatgcagag aggggagatg ggggacagtc tgaccctagg tgaacttgtt tattgcagct 3421 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca 3481 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc 3541 gggaattcgg cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta 3601 ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat accacatttg 3661 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa 3721 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca 3781 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 3841 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc 3901 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc 3961 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag 4021 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc 4081 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa 4141 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg 4201 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt 4261 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc 4321 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 4381 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 4441 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt 4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt 4561 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 4621 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 4681 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 4741 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 4801 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa 4861 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 4921 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 4981 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca 5041 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 5101 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 5161 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 5221 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 5281 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 5341 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc 5401 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 5461 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 5521 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 5581 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 5641 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag 5701 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg 5761 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg 5821 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa 5881 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt 5941 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg 6001 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca 6061 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt 6121 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga 6181 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca 6241 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac 6301 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg 6361 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag 6421 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac 6481 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt 6541 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg 6601 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc 6661 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc 6721 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc 6781 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa 6841 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt 6901 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat //