LOCUS       sequence-431771-        5628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses T-GECO1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5628)
  AUTHORS   Aggarwal A, Sunil S, Bendifallah I, Moon M, Drobizhev M, Zarowny L, 
            Zheng J, Wu SY, Lohman AW, Tebo AG, Emiliani V, Podgorski K, Shen Y,
            Campbell RE
  TITLE     Blue-shifted genetically encoded Ca(2+) indicator with enhanced 
            two-photon absorption.
  JOURNAL   Neurophotonics. 2024 Apr;11(2):024207. doi: 
            10.1117/1.NPh.11.2.024207.
  PUBMED    38577628
REFERENCE   2  (bases 1 to 5628)
  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: 
            "Neurophotonics."; date: "2024-04"; volume: "11(2):024207. doi"; 
            pages: " 10.1117/1.NPh.11.2.024207"
COMMENT     Sequence Label: pAAV hSyn RSET-T-GECO1.WPRE
FEATURES             Location/Qualifiers
     source          1..5628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    637..661
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      662..671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             680..697
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             701..733
                     /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(737..745,746..760)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1697..1753
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RHKWNKTGHAVRAIGRLSS"
     protein_bind    complement(2018..2042)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2052..2640
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2105..2125)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2523..2534)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2670..2687)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2676..2883
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2891..3031
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2891..3020
                     /label=AAV2 ITR
     rep_origin      3106..3561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3193..3212)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3403..3424
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3578..3597)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3697..3719
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3757..3775)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3843..3947
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3948..4016,4017..4808)
                     /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     complement(4166..4185)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgct gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagcaa gtttgtacaa aaaagcaggc
      661 tgccaccatg gggggttctc atcatcatca tcatcatggt atggctagca tgactggtgg
      721 acagcaaatg ggtcgggatc tgtacgacga tgacgataag gatccgagct cgagagtggt
      781 gagcaagggc gatgagacca caatgggcgt aatcaagccc gacatgaaga tcaagctgaa
      841 gatggagggc aacgtgaatg gccacgcctt cgtgatcgag ggcgagggcg agggcaagcc
      901 ctacgacggc acccacacca tcaacctgga ggtgaaggag ggagcccccc tgcccttctc
      961 ctacgacatc ctgaccaccg cgttcgccta cggcaacagg gccttcacca agtaccccga
     1021 cgacatcccc gactacttca agcagtcctt ccccgagggc tactcttggg agcgcatcat
     1081 gaccttcgag gacaagggca tcgtgaaggt gaagtccgac atctcgatgg aggagggctc
     1141 cttcatctac gaaatatacc tcaagggcga gaacttcccc cccaacggtc ccgtgatgca
     1201 gaagaagacc accggctggc ggaatcgtga tcaactgact gaagagcaga tcgcagaatt
     1261 taaagaggct ttctccctat ttgataagga cggggatgga acaataacaa ccgaggagct
     1321 ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc aggacatgat
     1381 caatgtagta gatgccgacg gtgacggcgc aatcgacttc cctgagttcc tgacaatgat
     1441 gtcaagaaaa atgaactaca gggacacgga agaagaaatt agagaagcgt tcggtgtgtt
     1501 tgataaggat ggcaatggct acatcagtgc agcagagctt caccgcgtga tgacaaacct
     1561 tggagaaaag ttatcagatg aagaggttga tgatgtgatc agggaagcag acatcgatgg
     1621 ggatggtctg gtaaactacg aagagtttgt acaaatgatg acagcgaagg gtggcggagg
     1681 ttctgtcgac tcatcacgtc ataagtggaa taagacaggt cacgcagtca gagctatagg
     1741 tcggctgagc tcaattgggg gtgagaagat gtacgtgcgc gacggcgtgc tgaagggcga
     1801 cgtcgagcac gagctgctgc tgaagggcgg cggccaccac cgcgttgact tcaagaccat
     1861 ctacagggcc aagaaggcgg tgaagctgcc cgactatcac tttgtggacc accacatcga
     1921 gatcctgaac cacgacaagg actacaacaa ggtgaccgtc tacgagagcg ccgcggcccg
     1981 caactcccgt aggggcatgg acgagctgta caagtaaacc cagctttctt gtacaaagtg
     2041 ggaattccga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2101 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2161 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2221 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2281 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2341 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2401 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     2461 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2521 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     2581 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     2641 atcgggaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2701 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2761 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2821 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     2881 ggagggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     2941 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     3001 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     3061 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     3121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3181 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3241 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3301 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     3361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3421 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     3481 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3541 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     3601 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     3661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     3721 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     3781 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     3841 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3901 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3961 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4021 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4081 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4141 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4201 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4261 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4321 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4381 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4441 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4501 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4561 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4621 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     4681 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4741 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4801 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
