LOCUS       sequence-445057-        7354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription for Oocyte expression of 1 TREK-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Deal PE, Lee H, Mondal A, Lolicato M, Mendonca PRF, Black H, Jang S,
            El-Hilali X, Bryant C, Isacoff EY, Renslo AR, Minor DL Jr
  TITLE     Development of covalent chemogenetic K(2P) channel activators.
  JOURNAL   Cell Chem Biol. 2024 Jul 18;31(7):1305-1323.e9. doi: 
            10.1016/j.chembiol.2024.06.006.
  PUBMED    39029456
REFERENCE   2  (bases 1 to 7354)
  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"; doi: 
            "10.1016/j.chembiol.2024.06"; journalName: "Cell Chem Biol"; date: 
            "2024-07-18- 18"; volume: "31"; issue: "7"; pages: "1305-1323.e9"
COMMENT     Sequence Label: pcDNA3.1 TREK-2
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1211..1230)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1402..1781
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1782..1985
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1936..1956
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2030..2049
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2030..2048
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2083..2125
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           3851..3976
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(4112..4129)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4118..4342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4388..4816
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4475..4494)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4685..4706
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4825..4845)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4830..5159
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5010..5145
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5072..5091
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5226..6020
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5280..5299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5890..5909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6194..6315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6231..6250)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6285..6304
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6364..6380)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6364..6380)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6377..6399)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6388..6404
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6412..6418,6419..6436,6437..6442))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6457..6478
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6595..6612)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6766..7354)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6846..6865)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     1381 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     1441 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1501 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1561 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     1621 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     1681 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     1741 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     1801 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     1861 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     1921 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     1981 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2041 actataggga gacccaagct ggctagctcg gtacccagct tgcttgttct ttttgcagaa
     2101 gctcagaata aacgctcaac tttggcagat caattccccg gggatccact agtccagtgt
     2161 ggtggaattc tgcagatatg aaatttccaa tcgagacgcc aagaaagcag gtgaactggg
     2221 atcctaaagt ggccgtgcct gcagcagccc caccagtgtg ccagcccaag agtgccacta
     2281 acgggcatca ccctgttcct cgactctcca tttcctctcg agccacggtg gtagccagga
     2341 tggaaggcgc ctcccaggga ggcctgcaaa cagttatgaa gtggaaaaca gtggttgcca
     2401 tcttcgtggt ggtcgtggtc tacctcgtca ctggtggcct tgtcttccgg gccttggaac
     2461 agcccttcga gagcagtcag aagaacacaa tcgccttgga gaaggcagaa ttcttgagag
     2521 atcacatctg tgtgagtccc caggaactag agacactgat tcagcatgca ctcgatgctg
     2581 ataacgcggg agtcagccca gtaggaaact cttccaacag cagcagtcac tgggaccttg
     2641 gaagtgcctt cttctttgct gggacagtca tcacaaccat agggtatggg aatattgctc
     2701 cgagcactga aggaggcaaa atcttttgta ttttatatgc catctttggg atcccgcttt
     2761 ttggtttctt attggctgga attggagacc aacttggaac catctttggc aaaagcattg
     2821 caagagtgga aaaggttttt cgaaagaagc aagtgagcca gaccaagatc cgggtcatct
     2881 ccacgattct cttcatcttg gctggctgca tcgtgtttgt gaccatccct gctgtcatct
     2941 ttaaatacat cgagggttgg accgccttgg agtccatcta ctttgtggta gtcacgctga
     3001 ccacagtagg ctttggtgat tttgtggcag ggggaaatgc tggcatcaat taccgagagt
     3061 ggtacaagcc gctggtgtgg ttttggatcc ttgttggcct tgcctacttt gcagctgtcc
     3121 tcagtatgat cggagactgg ctgcgggttt tatccaaaaa gacaaaagaa gaggttggtg
     3181 agatcaaagc ccacgcagct gagtggaagg ctaatgtcac tgctgagttc cgggagacga
     3241 gaaggcggct cagcgttgag atccatgaca agctgcaacg ggcagccact atccgaagta
     3301 tggagcgccg gaggctgggc ttggaccaga gggcccactc actggacatg ctatcccctg
     3361 agaagcgttc tgtctttgca gccctggaca caggccgttt caaggcctca tcgcaggaga
     3421 gtatcaacaa cagacccaac aacctacgcc ttaaggggcc agaacagctt accaaacatg
     3481 ggcagggcgc ttctgaggac aacatcatca acaagtttgg gtccacctcc aaactcacaa
     3541 agaggaaaaa caaagatctc aaaaagacct tacccgagga tgttcagaaa atctacaaaa
     3601 ccttccggaa ttactctctg gatgaagaga agaaagagga cgagacagaa aagatgtgta
     3661 actccgacaa ctccagcaca gccatgctga cagagtgtat acagcagcaa gctgagatgg
     3721 agaacggaat ggtacccacg gacaccaaag accaggggct ggagaacaat tctttacttg
     3781 aagacagaaa ctaaatccag cacagtggcg gccgctcgag tctagagcaa gcttgatctg
     3841 gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac ataataccaa
     3901 cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc tcctaataaa
     3961 aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaaaaaaaa annnncmccc
     4021 cccccccccc ctgcaatacg tacgcgtgcc cgggctagat ttaaatgtta acttaattaa
     4081 gctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     4141 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4201 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4261 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     4321 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     4381 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4441 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4501 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4561 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4621 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4681 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4741 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4801 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4861 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4921 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     4981 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5041 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5101 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5161 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5221 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5281 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5341 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5401 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5461 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5521 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5581 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5641 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5701 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5761 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5821 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5881 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5941 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6001 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6061 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6121 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6181 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6241 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6301 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6361 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6421 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6481 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6541 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6601 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6661 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6721 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6781 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6841 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6901 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6961 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7021 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7081 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7141 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7201 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     7261 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7321 gcagcagatt acgcgcagaa aaaaaggatc tcaa
//
