LOCUS       sequence-356929-        7299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TVMVP-inducible mCherry-conjugated Kir2.1 RELEASE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  AUTHORS   Vlahos AE, Kang J, Aldrete CA, Zhu R, Chong LS, Elowitz MB, Gao XJ
  TITLE     Protease-controlled secretion and display of intercellular signals.
  JOURNAL   Nat Commun. 2022 Feb 17;13(1):912. doi: 10.1038/s41467-022-28623-y.
  PUBMED    35177637
REFERENCE   2  (bases 1 to 7299)
  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"; journalName: "Nat 
            Commun."; date: "2022-02-17"; volume: "13(1):912. doi"; pages: " 
            10.1038/s41467-022-28623-y"
COMMENT     Sequence Label: CMVTO-mCherry-Kir2.1-HA-tvmvs-RXR-V2
FEATURES             Location/Qualifiers
     source          1..7299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        462..665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     616..636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    667..685
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    688..706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     716..740
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             844..1551
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(989..1007)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1257..1276
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      1624..1633
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1906..1932
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2887..2907
                     /codon_start=1
                     /product="tobacco vein mottling virus (TVMV) NIa protease 
                     recognition and cleavage site
                     "
                     /label=TVMV site
                     /translation="ETVRFQS"
     CDS             2979..3008
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3064..3081)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3070..3294
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3466..3485)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3617..3664
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3672..4694
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3672..4694
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4824..4945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4861..4880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4915..4934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4994..5010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4994..5010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5007..5029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5018..5034
                     /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(5042..5048,5049..5066,5067..5072))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5087..5108
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5225..5242)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5396..5984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5476..5495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6155..6946,6947..7015))
                     /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     6778..6797
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7016..7120)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7195..7214)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
       61 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      121 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      181 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      241 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      301 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      361 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      421 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      481 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      541 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      601 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      661 gagctctccc tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc
      721 gtttagtgaa ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa
      781 gacaccggga ccgatccagc ctccggactc tagcgtttaa acttaagctt cgccaccgct
      841 agcatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
      901 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      961 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     1021 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     1081 caccccgccg acattcccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     1141 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     1201 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     1261 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     1321 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     1381 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     1441 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     1501 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtctggcgcc
     1561 atgggcagtg tgcgaaccaa ccgctacagc atcgtctctt cagaagaaga cggtatgaag
     1621 ttggccacca tggcagttgc aaatggcttt gggaacggga agagtaaagt ccacacccga
     1681 caacagtgca ggagccgctt tgtgaagaaa gatggccact gtaatgttca gttcatcaat
     1741 gtgggtgaga aggggcaacg gtatctcgca gacatcttca ccacgtgtgt ggacattcgc
     1801 tggcggtgga tgctggttat cttctgcctg gctttcgtcc tgtcatggct gttttttggc
     1861 tgtgtgtttt ggttgatagc tctgctccat ggggacctgg atgcataccc atacgatgtt
     1921 ccagattacg cttccaaaga gggcaaggct tgtgtgtccg aggtcaacag cttcacggct
     1981 gccttcctct tctccattga gacccagaca accataggct atggtttcag atgtgtcacg
     2041 gatgaatgcc caattgctgt tttcatggtg gtgttccagt caatcgtggg ctgcatcata
     2101 gatgctttca tcattggcgc agtcatggcc aagatggcaa agccaaagaa gagaaacgag
     2161 actcttgtct tcagtcacaa tgccgtgatt gccatgagag acggcaagct gtgtttgatg
     2221 tggcgagtgg gcaatcttcg gaaaagccac ttggtggaag ctcatgttcg agcacagctc
     2281 ctcaaatcca gaattacttc tgaaggggag tatatccctc tggatcaaat agacatcaat
     2341 gttgggtttg acagtggaat agatcgtata tttctggtgt ccccaatcac tatagtccat
     2401 gaaatagatg aagacagtcc tttatatgat ttgagtaaac aggacattga caacgcagac
     2461 tttgaaatcg tggtcatact ggaaggcatg gtggaagcca ctgccatgac gacacagtgc
     2521 cgtagctctt atctagcaaa tgaaatcctg tggggccacc gctatgagcc tgtgctcttt
     2581 gaagagaagc actactacaa agtggactat tccaggttcc acaaaactta cgaagtcccc
     2641 aacactcccc tttgtagtgc cagagactta gcagaaaaga aatatatcct ctcaaatgca
     2701 aattcatttt gctatgaaaa tgaagttgcc ctcacaagca aagaggaaga cgacagtgaa
     2761 aatggagttc cagaaagcac tagtacggac acgccccctg acatagacct tcacaaccag
     2821 gcaagtgtac ctctagagcc caggccctta cggcgagagt cggagatact cgagggtggg
     2881 ggttcagaga cggttagatt ccagagtggg gggggcggct ctggtaccaa gcacatcctc
     2941 ttccgacgga ggagaagggg cttcaggcag tagggcccga acaaaaactc atctcagaag
     3001 aggatctgaa tagcgccgtc gaccatcatc ataatcatca ttgagtttaa acccgctgat
     3061 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3121 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3181 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3241 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggagatct
     3301 atcgatgttc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     3361 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3421 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3481 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3541 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3601 atggttcacg tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa
     3661 cttccttggc caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa
     3721 agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca
     3781 gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct
     3841 acaaagatcg ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc
     3901 ttgacattgg ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg
     3961 tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg
     4021 ccatggatgc gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac
     4081 cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc
     4141 atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc
     4201 tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg
     4261 atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga
     4321 gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt
     4381 ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag
     4441 gatcgccgcg gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct
     4501 tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc
     4561 gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga
     4621 ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga
     4681 gggcaaagga atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt
     4741 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4801 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4861 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4921 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4981 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     5041 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     5101 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     5161 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5221 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5281 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5341 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5401 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5461 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5521 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5581 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5641 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5701 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5761 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5821 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5881 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5941 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6001 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6061 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6121 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6181 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6241 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6301 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6361 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6421 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6481 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6541 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6601 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6661 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6721 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6781 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6841 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6901 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6961 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7021 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7081 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7141 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7201 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7261 tgagtagtgc gcgagcaaaa tttaagctac aacaaggca
//
