LOCUS       sequence-478760-        6441 bp    DNA     circular SYN 16-JUL-2025
DEFINITION  Red calcium indicator jRGECO1a with a nuclear export signal 
            expressed under a glial specific promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  TITLE     Fyhn unpublished plasmids
REFERENCE   2  (bases 1 to 6441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV gfaABC1D:NES-jRGECO1a
FEATURES             Location/Qualifiers
     source          1..6441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1158..1177
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1158..1176
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1249..1266
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1270..1302
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(1306..1314,1315..1329)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1357..1413
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKAGHAVRAIGRLSS"
     primer_bind     2109..2128
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2602..3190
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2655..2675)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3073..3084)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3208..3329
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3219..3238)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3273..3292
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3491..3620
                     /label=AAV2 ITR
     primer_bind     complement(3639..3656)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3639..3655)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3648..3670)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3797..4252
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3884..3903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4094..4115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4278..4382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4383..4451,4452..5243)
                     /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      5414..6002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5903..5922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6290..6311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6326..6331,6332..6349,6350..6356)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6364..6380
                     /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)."
     primer_bind     6369..6391
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6388..6404
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6388..6404
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct aacatatcct ggtgtggagt aggggacgct gctctgacag aggctcgggg
      241 gcctgagctg gctctgtgag ctggggagga ggcagacagc caggccttgt ctgcaagcag
      301 acctggcagc attgggctgg ccgcccccca gggcctcctc ttcatgccca gtgaatgact
      361 caccttggca cagacacaat gttcggggtg ggcacagtgc ctgcttcccg ccgcacccca
      421 gcccccctca aatgccttcc gagaagccca ttgagcaggg ggcttgcatt gcaccccagc
      481 ctgacagcct ggcatcttgg gataaaagca gcacagcccc ctaggggctg cccttgctgt
      541 gtggcgccac cggcggtgga gaacaaggct ctattcagcc tgtgcccagg aaaggggatc
      601 aggggatgcc caggcatgga cagtgggtgg caggggggga gaggagggct gtctgcttcc
      661 cagaagtcca aggacacaaa tgggtgaggg gagagctctc cccatagctg ggctgcggcc
      721 caaccccacc ccctcaggct atgccagggg gtgttgccag gggcacccgg gcatcgccag
      781 tctagcccac tccttcataa agccctcgca tcccaggagc gagcagagcc agagcaggtt
      841 ggagaggaga cgcatcacct ccgctgctcg caagctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtacttaa tacgactcac tatagggcta gcatgctgca gaacgagctt
     1201 gctcttaagt tggctggact tgatattaac aagactggag gaggttctca tcatcatcat
     1261 catcatggta tggctagcat gactggtgga cagcaaatgg gtcgggatct gtacgacgat
     1321 gacgataagg atctcgcaac aatggtcgac tcatcacgtc gtaagtggaa taaggcaggt
     1381 cacgcagtca gagctatagg tcggctgagc tcacccgtgg tttccgagcg gatgtacccc
     1441 gaggacggcg ccctgaagag cgagatcaag aaggggctga ggctgaagga cggcggccac
     1501 tacgccgccg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     1561 tacatcgtgg acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa
     1621 cagtgcgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga
     1681 ggtacaggcg ggagtctggt gagcaagggc gaggaggata acatggccat catcaaggag
     1741 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     1801 ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa ggtgaccaag
     1861 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1921 gcctacatta agcacccagc cgacatcccc gactacttca agctgtcctt ccccgagggc
     1981 ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt gaaccaggac
     2041 tcctccctgc aggacggcgt attcatctac aaggtgaagc tgcgcggcac caacttcccc
     2101 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggctacgcg tgacgacctg
     2161 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
     2221 gggacgataa caaccaagga gcttgggacg gtgttccgat cgctggggca gaaccccaca
     2281 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacattcgac
     2341 ttccctgagt tcctgacgat gatggcaaga aaaatgaatg acacagacag tgaagaggaa
     2401 attcgcgaag cgttccgcgt gtttgataag gacggcaatg gctacatcgg cgcagcagag
     2461 cttcgccacg tgatgacaga ccttggagag aagttaacag atgaggaggt tgatgaaatg
     2521 atcagggtag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg
     2581 atgacagcga agtgagaatt caatcaacct ctggattaca aaatttgtga aagattgact
     2641 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2701 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2761 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2821 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2881 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2941 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3001 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3061 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3121 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3181 gcctccccgc gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa
     3241 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3301 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3361 aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     3421 aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc
     3481 attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     3541 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3601 gtgagcgagc gagcgcgcag ccttaattaa cctaattcac tggccgtcgt tttacaacgt
     3661 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3721 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3781 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3841 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3901 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3961 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4021 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4081 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4141 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4201 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4261 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4381 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4501 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4561 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4621 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4681 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4801 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4921 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4981 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5221 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5521 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6001 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6061 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6181 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6241 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6301 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6361 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat
     6421 ttaattaagg ccttaattag g
//
