LOCUS       sequence-273253-        7002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  JxON-post.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7002)
  AUTHORS   Kim J, Zhao T, Petralia RS, Yu Y, Peng H, Myers E, Magee JC
  TITLE     mGRASP enables mapping mammalian synaptic connectivity with light 
            microscopy.
  JOURNAL   Nat Methods. 2011 Dec 4;9(1):96-102. doi: 10.1038/nmeth.1784.
  PUBMED    22138823
REFERENCE   2  (bases 1 to 7002)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-12-4- 4"; volume: "9"; 
            issue: "1"; pages: "96-102"
COMMENT     Sequence Label: paavCAG-Jx-rev-post-mGRASP-2A-dTomato
FEATURES             Location/Qualifiers
     source          1..7002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..278
                     /label=chicken beta-actin promoter
     primer_bind     557..579
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     641..660
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(692..725)
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1450..1503)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1900..1911
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    2980..3013
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3026..3614
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3079..3099)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3497..3508)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3657..3778
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3668..3687)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3722..3741
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4284..4872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4364..4383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5043..5834,5835..5903))
                     /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     5666..5685
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5904..6008)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6034..6489)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6171..6192)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6383..6402
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6561..6690
                     /label=AAV2 ITR
ORIGIN
        1 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
       61 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg gggggggggg
      121 ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg cggagaggtg
      181 cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc
      241 ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc
      301 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
      361 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg
      421 tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg
      481 gccctttgtg cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg
      541 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
      601 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
      661 ctgtctcatc attttggcaa agaattggat cataacttcg tatagcatac attatacgaa
      721 cggtaaagct tacttataca gctcatccat gccatacaag aacaggtgat gccgaccttc
      781 tgagcgttcg tactgctcga cgattgtgta gtcctcgttg tgactcgtga tgtccaactt
      841 ggtgtccacg tagtagtagc caggcagctg aaccggcttt ttggccatat atatagtttt
      901 gaactcaacc aggtagtgcc cgccatcttt gagtttaagc gcctgatgga tttcgccttt
      961 aagaacgcca tcgcgggggt acagtcgttc tgtagaggcc tcccacccca tggtcttctt
     1021 ctgcatcacg gggccatcgg ggggaaaatt tgttccgcgc atcttcacct tgtaaatcag
     1081 ggtgccgtcc tgcagagagg agtcttgcgt gacagtgacc agcccaccat cctcaaaatt
     1141 cattactcgt tcccacttaa atccttcagg aaatgacagt tttttgtaat caggtatatc
     1201 agcagggtgt ttgacgtaag ccttgctacc gtacatgaat tgagggctga ggatgtccca
     1261 ggcgaaagga agggggccac ctttggtaac cttgagcttg gcggtttgtg tgccctcata
     1321 tggtcttcct tcaccctcac cttcaatttc aaactcgtgc ccgttcatgc taccctccat
     1381 tctaactttg aatctcataa actccttaat tacttcctca cccttagaca ccattgtcga
     1441 ccccggggcc ggtccgggat tctcctcgac gtcgccacat gtgagcagag agccccgacc
     1501 ctcagatctg ccaccgctgc cacccacccg tgttgtgctg tgggagtggg ggtgtggatg
     1561 cggatgaggg agagtattat tctgtccacc agtgaatgtg ttgaaagtgt ggagaggctg
     1621 gattcctgga atagtgttgg gaatcatggt tatggtattg ggtgtcatca ggggaatgtc
     1681 gtctgggctt cggcgcattg ccagggtata gtccggaggg catgctgtcc tcagcaccac
     1741 ctcatgggga tggatggatt cacactcgtg gtccaggtct gtgtgtttca tctgaagact
     1801 catgatctcc tcctcaggtg cgtgcgtaag atcattggtt gtagttcgtt gcggactaca
     1861 tctcctatgt acgtcgtggc gccttttgtc cttcttgtaa tagagggccg cgaaagccag
     1921 aatgttcagg aaaagcaggg aagctcctac agctatggtc acggacagct ctgtgctgta
     1981 gtccctttgg tctacgctaa atggactagg ctgttgtttg gggtcgtctt gcttagcggt
     2041 tggaaatgcg ctggtcacag gggttgagtt tttccgagtt ggtcgaaggg tgatgtcagt
     2101 gcttgggacc ttggtggttg tacttgtgta ttggctgatg tcattgaggt tatgcaagtg
     2161 gggcaccagc tcgagcgtcc ctttctcatt agggtccttg gacaggactg tttgggtact
     2221 gaggtagtgg ttgtctggca ggagaacagg tccatcgcca atgggcgtgt tttgctggta
     2281 gtggtctgcc agctggacac tgccatcctc gacattatgt cttaccgtga aattggcctt
     2341 tatgccattc ttctgcttat ctgctgtaat atacacgttg tgactgttga agttgtactc
     2401 caatttgtgt cccaggatat ttccgtcctc cttaaaatcg gtgcctttga gctcgatgcg
     2461 gttaacgagt gtatcaccct cgaacttgac gactgctctt gtcttatatt tgccatcgtc
     2521 cttaaaagag atggtgcgct cctgaacgta accctcgggc atggcactct tgaagaagtc
     2581 atgcctcttc atatggtccg ggtaccgaga gaaacattgc accccgtagg tgagtgttgt
     2641 cacgagggtt ggccaaggca cgggaagttt tccagtggta cagatgaatt tcagcgtgag
     2701 tttcccgatg gtggcatcgc cttctccctc tcctctgaca gaaaacttat ggccattcac
     2761 gtcaccatcg agttccacga gaatagggac tacgccagta aacaactctt ctcccttgga
     2821 catgtcatcc agcttctgtg aaagaacgtg gaaagtctgg aggaggcaac ccaacaggca
     2881 gagtgtaagt ggggcgccac tccctcggtg caccacgcag gccatcatag ctcgccatac
     2941 ataattaggc cacatacatc taggaagtgc catggatcta taacttcgta taatgtatgc
     3001 tatacgaacg gtacagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3061 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3121 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3181 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3241 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3301 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3361 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3421 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     3481 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     3541 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     3601 ggccgcctcc ccgcatcgat accgtcgacc cgggcggccg cttcgagcag acatgataag
     3661 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     3721 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     3781 caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta
     3841 aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct
     3901 acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt
     3961 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgcccgggct ttgcccgggc
     4021 ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg cgcggggaga
     4081 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4141 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4201 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4261 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4321 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4381 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4441 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4501 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4561 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4621 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4681 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4741 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4801 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4861 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4921 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4981 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5041 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5101 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5161 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5221 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5281 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5341 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5401 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5461 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5521 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5581 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5641 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5701 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5761 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5821 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5881 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5941 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6001 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     6061 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6121 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6181 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6241 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6301 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6361 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6421 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6481 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6541 cctcttcgct attacgccag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
     6601 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     6661 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
     6721 tatctacgta gccatgctct aggaagatcg taccattgac gtcaataatg acgtatgttc
     6781 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6841 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6901 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6961 cttggcagta catctacgta ttagtcatcg ctattaccat gg
//
