LOCUS       sequence-461076-        6796 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  post.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6796)
  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 6796)
  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-post-mGRASP-2A-dTomato
FEATURES             Location/Qualifiers
     source          1..6796
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    968..1089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1005..1024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1059..1078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1132..1720
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1238..1249
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1647..1667
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2451..2504)
                     /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             2901..2912
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4007..4026)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4088..4110)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4389..4666)
                     /label=chicken beta-actin promoter
     repeat_region   4979..5108
                     /label=AAV2 ITR
     rep_origin      5180..5635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5267..5286)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5477..5498
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5661..5765
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5766..5834,5835..6626)
                     /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(5984..6003)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag cgagcgcgca gagagggagt
      781 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
      841 tacgtagcca tgctctagga agatccttat cgattttacc acatttgtag aggttttact
      901 tgctttaaaa aacctcccac atctccccct gaacctgaaa cataaaatga atgcaattgt
      961 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1021 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1081 tgtatcttat catgtctgct cgaagcggcc gcccgggtcg acggtatcga tgcggggagg
     1141 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
     1201 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
     1261 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
     1321 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
     1381 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
     1441 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
     1501 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
     1561 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
     1621 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
     1681 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt atcgataagc ttacttatac
     1741 agctcatcca tgccatacaa gaacaggtga tgccgacctt ctgagcgttc gtactgctcg
     1801 acgattgtgt agtcctcgtt gtgactcgtg atgtccaact tggtgtccac gtagtagtag
     1861 ccaggcagct gaaccggctt tttggccata tatatagttt tgaactcaac caggtagtgc
     1921 ccgccatctt tgagtttaag cgcctgatgg atttcgcctt taagaacgcc atcgcggggg
     1981 tacagtcgtt ctgtagaggc ctcccacccc atggtcttct tctgcatcac ggggccatcg
     2041 gggggaaaat ttgttccgcg catcttcacc ttgtaaatca gggtgccgtc ctgcagagag
     2101 gagtcttgcg tgacagtgac cagcccacca tcctcaaaat tcattactcg ttcccactta
     2161 aatccttcag gaaatgacag ttttttgtaa tcaggtatat cagcagggtg tttgacgtaa
     2221 gccttgctac cgtacatgaa ttgagggctg aggatgtccc aggcgaaagg aagggggcca
     2281 cctttggtaa ccttgagctt ggcggtttgt gtgccctcat atggtcttcc ttcaccctca
     2341 ccttcaattt caaactcgtg cccgttcatg ctaccctcca ttctaacttt gaatctcata
     2401 aactccttaa ttacttcctc acccttagac accattgtcg accccggggc cggtccggga
     2461 ttctcctcga cgtcgccaca tgtgagcaga gagccccgac cctcagatct gccaccgctg
     2521 ccacccaccc gtgttgtgct gtgggagtgg gggtgtggat gcggatgagg gagagtatta
     2581 ttctgtccac cagtgaatgt gttgaaagtg tggagaggct ggattcctgg aatagtgttg
     2641 ggaatcatgg ttatggtatt gggtgtcatc aggggaatgt cgtctgggct tcggcgcatt
     2701 gccagggtat agtccggagg gcatgctgtc ctcagcacca cctcatgggg atggatggat
     2761 tcacactcgt ggtccaggtc tgtgtgtttc atctgaagac tcatgatctc ctcctcaggt
     2821 gcgtgcgtaa gatcattggt tgtagttcgt tgcggactac atctcctatg tacgtcgtgg
     2881 cgccttttgt ccttcttgta atagagggcc gcgaaagcca gaatgttcag gaaaagcagg
     2941 gaagctccta cagctatggt cacggacagc tctgtgctgt agtccctttg gtctacgcta
     3001 aatggactag gctgttgttt ggggtcgtct tgcttagcgg ttggaaatgc gctggtcaca
     3061 ggggttgagt ttttccgagt tggtcgaagg gtgatgtcag tgcttgggac cttggtggtt
     3121 gtacttgtgt attggctgat gtcattgagg ttatgcaagt ggggcaccag ctcgagcgtc
     3181 cctttctcat tagggtcctt ggacaggact gtttgggtac tgaggtagtg gttgtctggc
     3241 aggagaacag gtccatcgcc aatgggcgtg ttttgctggt agtggtctgc cagctggaca
     3301 ctgccatcct cgacattatg tcttaccgtg aaattggcct ttatgccatt cttctgctta
     3361 tctgctgtaa tatacacgtt gtgactgttg aagttgtact ccaatttgtg tcccaggata
     3421 tttccgtcct ccttaaaatc ggtgcctttg agctcgatgc ggttaacgag tgtatcaccc
     3481 tcgaacttga cgactgctct tgtcttatat ttgccatcgt ccttaaaaga gatggtgcgc
     3541 tcctgaacgt aaccctcggg catggcactc ttgaagaagt catgcctctt catatggtcc
     3601 gggtaccgag agaaacattg caccccgtag gtgagtgttg tcacgagggt tggccaaggc
     3661 acgggaagtt ttccagtggt acagatgaat ttcagcgtga gtttcccgat ggtggcatcg
     3721 ccttctccct ctcctctgac agaaaactta tggccattca cgtcaccatc gagttccacg
     3781 agaataggga ctacgccagt aaacaactct tctcccttgg acatgtcatc cagcttctgt
     3841 gaaagaacgt ggaaagtctg gaggaggcaa cccaacaggc agagtgtaag tggggcgcca
     3901 ctccctcggt gcaccacgca ggccatcata gctcgccata cataattagg ccacatacat
     3961 ctaggaagtg ccatggatcc aattctttgc caaaatgatg agacagcaca ataaccagca
     4021 cgttgcccag gagctgtagg aaaaagaaga aggcatgaac atggttagca gaggctctag
     4081 agccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc cgaaggcagc
     4141 cgtccccccg cggacagccc cgagccgctc cccccgcaca aagggccctc ccggagcccc
     4201 tcaaggcttt cacgcagcca cagaaaagaa acaagccgtc attaaaccaa gcgctaatta
     4261 cagcccggag gagaagggcc gtcccgcccg ctcacctgtg ggagtaacgc ggtcagtcag
     4321 agccggggcg ggcggcgcga ggcggcggcg gagcggggca cggggcgaag gcagcgcgca
     4381 gcgactcccg cccgccgcgc gcttcgcttt ttatagggcc gccgccgccg ccgcctcgcc
     4441 ataaaaggaa actttcggag cgcgccgctc tgattggctg ccgccgcacc tctccgcctc
     4501 gccccgcccc gcccctcgcc ccgccccgcc ccgcctggcg cgcgcccccc cccccccccc
     4561 gcccccatcg ctgcacaaaa taattaaaaa ataaataaat acaaaattgg gggtggggag
     4621 gggggggaga tggggagagt gaagcagaac gtggggctca cctcgaccat ggtaatagcg
     4681 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     4741 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     4801 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     4861 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatggtacg
     4921 atcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag
     4981 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     5041 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5101 gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5161 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5221 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5281 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5341 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5401 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5461 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5521 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5581 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     5641 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5701 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5761 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5821 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5881 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5941 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6001 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6061 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6121 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6181 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6241 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6301 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6361 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6421 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6481 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6541 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6601 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6661 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6781 agatcaaagg atcttc
//
