LOCUS       sequence-203927-        6239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Kir2.1.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CMV-GFP-Kir2.1
FEATURES             Location/Qualifiers
     source          1..6239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(657..659,660..662,663..1373)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(657..659,660..662,663..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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(join(5179..5970,5971..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
//
