LOCUS       sequence-243949-        6335 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mouse Connexin 36-GCaMP3 fusion for measuring gap junction localized
            calcium signaling - expresses in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6335)
  AUTHORS   Moore KB, Mitchell CK, Lin YP, Lee YH, Shihabeddin E, O'Brien J
  TITLE     Localized calcium signaling and the control of coupling at Cx36 gap 
            junctions.
  JOURNAL   eNeuro. 2020 Mar 11. pii: ENEURO.0445-19.2020. doi: 
            10.1523/ENEURO.0445-19.2020.
  PUBMED    32179580
REFERENCE   2  (bases 1 to 6335)
  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: "eNeuro. 
            2020 Mar 11. pii: ENEURO.0445-19.2020. doi: 
            10.1523/ENEURO.0445-19.2020."
COMMENT     Sequence Label: Cx36-GCaMP
FEATURES             Location/Qualifiers
     source          1..6335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1640..1648,1649..1666,1667..1669,1670..1702,1703..1705,
                     1706..1729,1730..1756,1757..1813,1814..1819,1820..2089,
                     2090..2107,2108..2542,2543..2545,2546..2548,2549..2992)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1640..1648,1649..1666,1667..1669,1670..1702,1703..1705,
                     1706..1729,1730..1756,1757..1813,1814..1819,1820..2089,
                     2090..2107,2108..2542,2543..2545,2546..2548,2549..2992)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNTRDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1736..1745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2026..2047
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2153..2174)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2414..2433)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3114..3235
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3151..3170)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3205..3224
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3242..3697)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3379..3400)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3591..3610
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3724..3828
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3830..4187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3830..4021
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3853..3873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4038..4173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4222..5016
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4222..5016
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4276..4295)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4886..4905
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5204..5223)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5248..5295
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5624..6212
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6113..6132
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcggg gaggggccag
      601 gaccaagaac gtgcccggta ctgcccagtc tttgtctgct gcctccggat gcacagcgat
      661 gggggaatgg accatcttgg agaggctgct ggaagccgcg gtgcagcagc actccactat
      721 gattgggagg atcctgttga ctgtggtggt gatcttccgg atactcattg tggccattgt
      781 aggggagacg gtgtacgatg atgagcagac catgtttgtg tgcaacaccc tacagcccgg
      841 ctgtaaccag gcctgctatg accgcgcctt tcccatctcc catatacgtt actgggtctt
      901 ccagatcata atggtgtgca cccccagtct ctgttttatc acctattctg tgcaccaatc
      961 cgccaagcag cgagaacgcc ggtactctac tgtcttccta gccctggaca gagaccctgc
     1021 tgagtctata gggggacctg gaggaactgg gggtgggggc agcggaggga gcaaacgaga
     1081 agataagaag ttgcaaaatg ccattgtcaa tggggtgctg cagaacacag agaccaccag
     1141 taaggagaca gaaccagatt gcttagaggt taaagagctg actccacatc catctgggct
     1201 gcgcacagca gcaaggtcca agctccgaag acaggaaggt atctcccgct tctacatcat
     1261 ccaagtggtg tttcgaaatg ctctggagat tgggtttctg gtgggccagt actttctata
     1321 tggcttcagt gttccagggt tgtatgagtg caaccgttac ccctgcatca aggaggtaga
     1381 atgttatgtg tctagaccta ccgagaagac agtctttctg gtgttcatgt ttgctgtgag
     1441 cggcatttgt gtggtgctca atctggctga acttaaccat ctgggatggc ggaagatcaa
     1501 actggctgtc cggggagccc aggccaagag gaagtcagtc tatgagatac gtaacaaaga
     1561 tctgcctcga gtcagtgttc ccaatttcgg caggactcag tccagtgact ctgcctatgt
     1621 gtgtgatctc gccaccatga tgggttctca tcatcatcat catcatggta tggctagcat
     1681 gactggtgga cagcaaatgg gtcgggatct gtacgacgat gacgataagg atctcgccac
     1741 catggtcgac tcatcacgtc gtaagtggaa taagacaggt cacgcagtca gagctatagg
     1801 tcggctgagc tcactcgaga acgtctatat caaggccgac aagcagaaga acggcatcaa
     1861 ggcgaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg cctaccacta
     1921 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1981 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     2041 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     2101 agggagcatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2161 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg agggcgatgc
     2221 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2281 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     2341 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     2401 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2461 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2521 ggggcacaag ctggagtaca acacgcgtga ccaactgact gaagagcaga tcgcagaatt
     2581 taaagaggct ttctccctat ttgacaagga cggggatggg acaataacaa ccaaggagct
     2641 ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat
     2701 caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc tgacaatgat
     2761 ggcaagaaaa atgaaagaca cagacagtga agaagaaatt agagaagcgt tccgtgtgtt
     2821 tgataaggat ggcaatggct acatcagtgc agcagagctt cgccacgtga tgacaaacct
     2881 tggagagaag ttaacagatg aagaggttga tgaaatgatc agggaagcag acatcgatgg
     2941 ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagt gagcggccgc
     3001 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3061 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3121 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3181 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3241 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3301 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3361 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3421 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3481 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3541 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3601 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3661 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3721 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3781 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3841 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4021 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4081 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4141 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4201 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4261 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4321 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4381 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4441 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4501 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4561 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4621 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4681 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4741 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4801 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4861 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4921 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4981 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5041 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5101 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5161 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5221 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5281 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5341 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5401 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5461 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5521 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5581 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5641 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5701 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5761 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5821 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5881 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5941 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6001 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6061 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6121 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6181 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6241 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6301 tggataaccg tattaccgcc atgcattagt tatta
//
