pHybrid-cpGFP-Kir3.2(mouse)-mScarlet

LOCUS       pHybrid-cpGFP-Kir3.2(mouse)-mScarlet               6281 bp DNA     circular SYN 19-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pCI-sodium:proton antiporter Bacillus sp. YZJH907-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6281)
  AUTHORS   admin
  TITLE     Direct Submission
  JOURNAL   Exported Apr 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        754..872
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             913..930
                     /codon_start=1
                     /label=6x-HisTag
                     /translation="HHHHHH"
     primer_bind     950..992
                     /label=V-CpGFP-R
     regulatory      961..966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             970..1692
                     /codon_start=1
                     /label=特征 1
                     /translation="NVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYN"
     CDS             970..1692
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="NVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPV
                     LLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYN"
     primer_bind     complement(1672..1701)
                     /label=cpGFP-R2
     primer_bind     1682..1720
                     /label=Kir-zebra-F2
     CDS             1693..1701
                     /codon_start=1
                     /label=Link
                     /translation="GTG"
     misc_feature    1702..2352
                     /label=Kir3.2-cyto-mouse
     primer_bind     complement(2333..2377)
                     /label=Kir-mouse-R
     primer_bind     2353..2380
                     /label=rrnB-F
     terminator      2365..2459
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     polyA_signal    2483..2604
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        2619..2863
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2714..2849
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2864..2869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2870..3565
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     polyA_signal    3570..3617
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3697..4152
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4484..4588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4589..5449
                     /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"
     rep_origin      5620..6208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tcgtttagtg aaccgtcaga tcactagaag cttcaccaca attcagcaaa ttgtgaacat
      781 catcacgttc atctttccct ggttgccaat ggcccatttt cctgtcagta acgagaaggt
      841 cgcgaattca ggcgcttttt agactggtcg taacccgttt ttggctaaca ggaggaatta
      901 accatgggga gccaccatca ccatcaccat gctagcctcg agaattcacg cgtggtacca
      961 gccaccatga acgtctatat caaggccgac aagcagaaga acggcatcaa ggcgaacttc
     1021 aagatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta ccagcagaac
     1081 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cgtgcagtcc
     1141 aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1201 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
     1261 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1321 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc cacctacggc
     1381 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1441 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1501 cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac catcttcttc
     1561 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1621 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1681 ctggagtaca acggcaccgg caagaagaga gccgagaccc tggtgttcag cacccacgcc
     1741 gtgatcagca tgagagacgg aaagctgtgc ctgatgttcc gggtgggaga cctgagaaac
     1801 agccacatcg tggaggccag catcagagcc aagctgatca agagcaaaca gaccagcgag
     1861 ggcgaattca ttccactgaa ccagaccgac atcaacgtgg gctattacac cggcgacgac
     1921 agactgttcc tggtgagccc cctgatcatc tcccacgaaa tcaaccagca gtcacccttc
     1981 tgggagatct ccaaagccca gctgcccaag gaagagctgg agatcgtggt gatcctggag
     2041 ggcatggtgg aggccaccgg catgacctgc caggccagaa gcagctacat cacaagtgag
     2101 atcctgtggg gatacagatt cacccccgtg ctgaccctgg aggatggctt ctatgaggtg
     2161 gactataatt ccttccatga gacatacgag accagcaccc ccagcctgag cgccaaagag
     2221 ctggccgaac tggccaacag agccgagctg cccctgagct ggagcgtgag cagcaagctg
     2281 aaccagcacg ccgaactgga gaccgaggag gaagaaaaaa accccgagga gctgaccgag
     2341 agaaacggct agtctagagt cgacccaggc atcaaataaa acgaaaggct cagtcgaaag
     2401 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatg
     2461 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     2521 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     2581 attataagct gcaataaaca agttaacaac aacaattggg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2701 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     2761 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     2821 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagccacca tggtgagcaa
     2881 gggcgaggca gtgatcaagg agttcatgcg gttcaaggtg cacatggagg gctccatgaa
     2941 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     3001 cgccaagctg aaggtgacca agggtggccc cctgcccttc tcctgggaca tcctgtcccc
     3061 tcagttcatg tacggctcca gggccttcac caagcacccc gccgacatcc ccgactacta
     3121 taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     3181 cgccgtgacc gtgacccagg acacctccct ggaggacggc accctgatct acaaggtgaa
     3241 gctccgcggc accaacttcc ctcctgacgg ccccgtaatg cagaagaaga caatgggctg
     3301 ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg ctgaagggcg acattaagat
     3361 ggccctgcgc ctgaaggacg gcggccgcta cctggcggac ttcaagacca cctacaaggc
     3421 caagaagccc gtgcagatgc ccggcgccta caacgtcgac cgcaagttgg acatcacctc
     3481 ccacaacgag gactacaccg tggtggaaca gtacgaacgc tccgagggcc gccactccac
     3541 cggcggcatg gacgagctgt acaagtgacc ggcaataaaa agacagaata aaacgcacgg
     3601 tgttgggtcg tttgttcgga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc
     3661 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     3721 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3781 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3841 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3901 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3961 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4021 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     4081 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4141 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4201 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4261 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4321 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4381 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4441 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4501 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4561 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4621 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4681 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4741 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4801 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4861 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4921 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4981 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5041 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5101 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5161 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5221 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5281 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5341 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5401 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5461 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5521 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5581 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5641 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5701 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5761 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5821 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5881 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5941 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6001 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6061 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6121 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6181 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6241 ggccttttgc tggccttttg ctcacatggc tcgacagatc t
//