LOCUS Exported 6239 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mammalian expression of Kir2.1. ACCESSION . VERSION . KEYWORDS CMV-GFP-Kir2.1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6239) TITLE Bikash Pattnaik Lab plasmids REFERENCE 2 (bases 1 to 6239) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6239 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 17..37 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" enhancer 98..401 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 402..605 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" protein_bind 626..650 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 651..660 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 657..1373 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 657..1373 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(702..723) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(963..982) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1310..1331 /label=EGFP-C /note="EGFP, forward primer" regulatory 1443..1452 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2670..2694) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" polyA_signal 2723..2844 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2734..2753) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2788..2807 /label=SV40pA-R /note="SV40 polyA, reverse primer" enhancer 2942..3321 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3322..3525 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" regulatory 3552..3561 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3558..3956 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" primer_bind complement(3994..4011) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4000..4224 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(4249..4266) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4420..5008) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4500..4519) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5179..6039) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5802..5821 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6040..6144) /gene="bla" /label=AmpR promoter primer_bind 6212..6230 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 ggcgcgccgc ggccgccaac tttgtataga aaagttgtag ttattaatag taatcaatta 61 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 121 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 181 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 241 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 301 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 361 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 421 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 481 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 541 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 601 gagctggttt agtgaaccgt cagatccaag tttgtacaaa aaagcaggct gccaccatgg 661 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg 721 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca 781 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg 841 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc 901 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca 961 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga 1021 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc 1081 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca 1141 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc 1201 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc 1261 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc 1321 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggaattcc 1381 ccggcagtgt gcgaaccaac cgctacagca tcgtctcttc agaagaagac ggtatgaagt 1441 tggccaccat ggcagttgca aatggctttg ggaacgggaa gagtaaagtc cacacccgac 1501 aacagtgcag gagccgcttt gtgaagaaag atggccactg taatgttcag ttcatcaatg 1561 tgggtgagaa ggggcaacgg tacctcgcag acatcttcac cacgtgtgtg gacattcgct 1621 ggcggtggat gctggttatc ttctgcctgg ctttcgtcct gtcatggctg ttttttggct 1681 gtgtgttttg gttgatagct ctgctccatg gggacctgga tgcatccaaa gagggcaaag 1741 cttgtgtgtc cgaggtcaac agcttcacgg ctgccttcct cttctccatt gagacccaga 1801 caaccatagg ctatggtttc agatgtgtca cggatgaatg cccaattgct gttttcatgg 1861 tggtgttcca gtcaatcgtg ggctgcatca tcgatgcttt catcattggc gcagtcatgg 1921 ccaagatggc aaagccaaag aagagaaacg agactcttgt cttcagtcac aatgccgtga 1981 ttgccatgag agacggcaag ctgtgtttga tgtggcgagt gggcaatctt cggaaaagcc 2041 acttggtgga agctcatgtt cgagcacagc tcctcaaatc cagaattact tctgaagggg 2101 agtatatccc tctggatcaa atagacatca atgttgggtt tgacagtgga atcgatcgta 2161 tatttctggt gtccccaatc actatagtcc atgaaataga tgaagacagt cctttatatg 2221 atttgagtaa acaggacatt gacaacgcag actttgaaat cgtggtcata ctggaaggca 2281 tggtggaagc cactgccatg acgacacagt gccgtagctc ttatctagca aatgaaatcc 2341 tgtggggcca ccgctatgag cctgtgctct ttgaagagaa gcactactac aaagtggact 2401 attccaggtt ccacaaaact tacgaagtcc ccaacactcc cctttgtagt gccagagact 2461 tagcagaaaa gaaatatatc ctctcaaatg caaattcatt ttgctatgaa aatgaagttg 2521 ccctcacaag caaagaggaa gacgacagtg aaaatggagt tccagaaagc actagtacgg 2581 acacgccccc tgacatagac cttcacaacc aggcaagtgt acctctagag cccaggccct 2641 tacggcgaga gtcggagata tgaggatcca cccagctttc ttgtacaaag tggtgatggc 2701 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga 2761 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc 2821 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt 2881 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtacgcgt 2941 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc 3001 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc 3061 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg 3121 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat 3181 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc 3241 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta 3301 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag 3361 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt 3421 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa 3481 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga 3541 gaacccactg cgccaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa 3601 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag 3661 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac 3721 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt 3781 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac 3841 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc 3901 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaactcg 3961 agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc 4021 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg 4081 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc 4141 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg 4201 ctggggatgc ggtgggctct atgggcggcc gcggcgctct tccgcttcct cgctcactga 4261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 4321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 4381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 4441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 4501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 4561 gcttaccgga tacctgtccg cctttctctc ttcgggaagc gtggcgcttt ctcatagctc 4621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 4681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 4741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 4801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 4861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 4921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 4981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 5521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 5581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 5641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 5701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 5761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 5821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 5881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 5941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga 6181 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc //