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    1..21
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    610..634
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      635..644
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             641..1357
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             641..1357
                     /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(686..707)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(947..966)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1294..1315
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1427..1436
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2654..2678)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    2707..2828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2718..2737)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2772..2791
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     enhancer        2926..3305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3306..3509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3536..3545
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3542..3940
                     /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(3978..3995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3984..4208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4233..4250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4404..4992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4484..4503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5163..6023)
                     /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     5786..5805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6024..6128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6196..6214
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caactttgta tagaaaagtt gtagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc caagtttgta caaaaaagca ggctgccacc atggtgagca agggcgagga
      661 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      721 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      781 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      841 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      901 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      961 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1021 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1081 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1141 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1201 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1261 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1321 cgccgggatc actctcggca tggacgagct gtacaaggaa ttccccggca gtgtgcgaac
     1381 caaccgctac agcatcgtct cttcagaaga agacggtatg aagttggcca ccatggcagt
     1441 tgcaaatggc tttgggaacg ggaagagtaa agtccacacc cgacaacagt gcaggagccg
     1501 ctttgtgaag aaagatggcc actgtaatgt tcagttcatc aatgtgggtg agaaggggca
     1561 acggtacctc gcagacatct tcaccacgtg tgtggacatt cgctggcggt ggatgctggt
     1621 tatcttctgc ctggctttcg tcctgtcatg gctgtttttt ggctgtgtgt tttggttgat
     1681 agctctgctc catggggacc tggatgcatc caaagagggc aaagcttgtg tgtccgaggt
     1741 caacagcttc acggctgcct tcctcttctc cattgagacc cagacaacca taggctatgg
     1801 tttcagatgt gtcacggatg aatgcccaat tgctgttttc atggtggtgt tccagtcaat
     1861 cgtgggctgc atcatcgatg ctttcatcat tggcgcagtc atggccaaga tggcaaagcc
     1921 aaagaagaga aacgagactc ttgtcttcag tcacaatgcc gtgattgcca tgagagacgg
     1981 caagctgtgt ttgatgtggc gagtgggcaa tcttcggaaa agccacttgg tggaagctca
     2041 tgttcgagca cagctcctca aatccagaat tacttctgaa ggggagtata tccctctgga
     2101 tcaaatagac atcaatgttg ggtttgacag tggaatcgat cgtatatttc tggtgtcccc
     2161 aatcactata gtccatgaaa tagatgaaga cagtccttta tatgatttga gtaaacagga
     2221 cattgacaac gcagactttg aaatcgtggt catactggaa ggcatggtgg aagccactgc
     2281 catgacgaca cagtgccgta gctcttatct agcaaatgaa atcctgtggg gccaccgcta
     2341 tgagcctgtg ctctttgaag agaagcacta ctacaaagtg gactattcca ggttccacaa
     2401 aacttacgaa gtccccaaca ctcccctttg tagtgccaga gacttagcag aaaagaaata
     2461 tatcctctca aatgcaaatt cattttgcta tgaaaatgaa gttgccctca caagcaaaga
     2521 ggaagacgac agtgaaaatg gagttccaga aagcactagt acggacacgc cccctgacat
     2581 agaccttcac aaccaggcaa gtgtacctct agagcccagg cccttacggc gagagtcgga
     2641 gatatgagga tccacccagc tttcttgtac aaagtggtga tggccggccg cttcgagcag
     2701 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2761 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2821 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2881 aggtttttta aagcaagtaa aacctctaca aatgtggtac gcgttgacat tgattattga
     2941 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     3001 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     3061 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     3121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     3181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     3241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     3301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     3361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     3421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     3481 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcgccac
     3541 catggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     3601 caacagcatc cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg
     3661 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     3721 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     3781 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     3841 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     3901 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa ctcgagtcta gagggcccgt
     3961 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4021 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4081 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4141 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4201 ctctatgggc ggccgcggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4321 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4381 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4441 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4501 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4561 tccgcctttc tctcttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4621 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4681 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4741 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4801 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4861 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4921 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4981 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5041 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5101 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5161 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5221 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5281 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5341 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5401 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5461 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5521 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5581 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5641 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5701 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5761 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5821 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5881 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5941 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6001 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6061 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6121 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6181 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcggcgcgc cgcggccgc
//
