LOCUS       sequence-461077-        6415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pre.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6415)
  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 6415)
  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-pre-mGRASP-mCerulean
FEATURES             Location/Qualifiers
     source          1..6415
                     /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    1095..1216
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1132..1151)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1186..1205
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    1259..1847
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1365..1376
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1774..1794
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3453..3500)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     primer_bind     complement(3626..3645)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3707..3729)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4008..4285)
                     /label=chicken beta-actin promoter
     repeat_region   4598..4727
                     /label=AAV2 ITR
     rep_origin      4799..5254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4886..4905)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5096..5117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5280..5384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5385..5453,5454..6245)
                     /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(5603..5622)
                     /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 gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg cgctcgctcg
      841 ctcactgagg ccgcccgggc aaagcccggg cggcctcagt gagcgagcga gcgcgcagag
      901 agggagtggc caactccatc actaggggtt ccttgtagtt aatgattaac ccgccatgct
      961 acttatctac gtagccatgc tctaggaaga tccttatcga ttttaccaca tttgtagagg
     1021 ttttacttgc tttaaaaaac ctcccacatc tccccctgaa cctgaaacat aaaatgaatg
     1081 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1141 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1201 tcatcaatgt atcttatcat gtctgctcga agcggccgcc cgggtcgacg gtatcgatgc
     1261 ggggaggcgg cccaaaggga gatccgactc gtctgagggc gaaggcgaag acgcggaaga
     1321 ggccgcagag ccggcagcag gccgcgggaa ggaaggtccg ctggattgag ggccgaaggg
     1381 acgtagcaga aggacgtccc gcgcagaatc caggtggcaa cacaggcgag cagccaagga
     1441 aaggacgatg atttccccga caacaccacg gaattgtcag tgcccaacag ccgagcccct
     1501 gtccagcagc gggcaaggca ggcggcgatg agttccgccg tggcaatagg gagggggaaa
     1561 gcgaaagtcc cggaaaggag ctgacaggtg gtggcaatgc cccaaccagt gggggttgcg
     1621 tcagcaaaca cagtgcacac cacgccacgt tgcctgacaa cgggccacaa ctcctcataa
     1681 agagacagca accaggattt atacaaggag gagaaaatga aagccatacg ggaagcaata
     1741 gcatgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc aacatagtta
     1801 agaataccag tcaatctttc acaaattttg taatccagag gttgattatc gataagcttg
     1861 atgtcaaact ctcacttgta cagctcatcc atacccaggg ttatgccggc agctgtgaca
     1921 aactccagca gtaccatatg atctctcttt tcgttggggt ccttgctcag cttgctctgt
     1981 gtactaaggt agtgattgtc gggcaacagc acaggtccat caccgatggg agtattctgt
     2041 tggtaatggt cagccagctg aacgcttcca tcctctatgt tgtgccggat tttgaaattg
     2101 gcctttatgc catttttctg cttgtcggcg gttatgtaaa cattatcaga tatggcgttg
     2161 tattccagct tgtggccgag aatgttccca tcttctttga agtcgattcc cttgagttca
     2221 atcctgttca ccaaggtgtc accttcgaat ttcacctccg cccttgtctt gtagttacca
     2281 tcgtccttga aaaatatcgt cctttcctga acatacccct cgggcatagc ggacttgaag
     2341 aagtcgtgct gcttcatgtg gtcagggtac cgggcgaagc actgtacacc ccatgtgagg
     2401 gtggtgacga gtgtgggcca aggcacgggg agcttccctg ttgtacagat gaacttcagc
     2461 gtcagtttgc cgtaggtggc atccccttct ccctcgccgc ttacggagaa cttgtgccca
     2521 tttacatctc cgtcgagctc caccaggata ggcacgaccc ctgtaaacaa ctcttcgccc
     2581 ttgctcacca tagatcttac gtagtactct ttatccttgt ttttcttgtt tttgtttgct
     2641 gatttggcgc tggagggctg tttctcttta accacggcgc cattactctg agcgctgtta
     2701 ctgatatagt tcctagattc gtcaacgtgg taagatccct catccctgtt gcgatacttg
     2761 tagcgtacgc agaagaagat gcccaggccg ataaacagaa gcaggccggc gactcccccg
     2821 agtacgatca gggccatcgg ctgtactggt gttgaccaag taggaagcac cttgatgttg
     2881 ctctccagca gcacctgtcc agagtcgcta agaagacact gccacatccc tgcctccggg
     2941 ttaaggaccc acacagcctt ctctctttta ctgactttag cctccttatt ctccagcttc
     3001 agagacagca tcagtttagg tgaggtggga ccccacactt cgcaggtcag gttcttctgc
     3061 agctgtgttg ccctcatcac cacgaggttg acctcctgat gcaactttcc ggtcttagcc
     3121 tccaaggcga gggtgaggtt cccggagcca gcatactggg gcagggcttg ggggagtgtc
     3181 aagtgcaaag gcaacttctt gcccatctgc agcttagggt cctgtgtgac tctcttcacg
     3241 ctcacttctt tgtttttcag atcaaaagtg atccaactct tgctggagga ggctctttcg
     3301 gcctgccacc acaactctcc tgatcctgtg agcttctcga ctgtgaaggc caatgggaaa
     3361 ctgaactcca cctgttcccc ctccttctta taaacaatag agctagcttt ctgaaagtcg
     3421 acgctaccac cgccgccaga tccgccaccg cctgtgatgc cagcagcgtt cacatattcg
     3481 tgcagcacca tgtggtcgcg ggaagcggtg acttctcctg tcttccaatc tgaagcggga
     3541 agagcgaagg ggagaagtgc agcaagcaac aaaagactag tagaaggtgg catggatcca
     3601 attctttgcc aaaatgatga gacagcacaa taaccagcac gttgcccagg agctgtagga
     3661 aaaagaagaa ggcatgaaca tggttagcag aggctctaga gccgccggtc acacgccaga
     3721 agccgaaccc cgccctgccc cgtccccccc gaaggcagcc gtccccccgc ggacagcccc
     3781 gagccgctcc ccccgcacaa agggccctcc cggagcccct caaggctttc acgcagccac
     3841 agaaaagaaa caagccgtca ttaaaccaag cgctaattac agcccggagg agaagggccg
     3901 tcccgcccgc tcacctgtgg gagtaacgcg gtcagtcaga gccggggcgg gcggcgcgag
     3961 gcggcggcgg agcggggcac ggggcgaagg cagcgcgcag cgactcccgc ccgccgcgcg
     4021 cttcgctttt tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa ctttcggagc
     4081 gcgccgctct gattggctgc cgccgcacct ctccgcctcg ccccgccccg cccctcgccc
     4141 cgccccgccc cgcctggcgc gcgccccccc cccccccccg cccccatcgc tgcacaaaat
     4201 aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat ggggagagtg
     4261 aagcagaacg tggggctcac ctcgaccatg gtaatagcga tgactaatac gtagatgtac
     4321 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     4381 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     4441 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     4501 actatgggaa catacgtcat tattgacgtc aatggtacga tcttcctaga gcatggctac
     4561 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     4621 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     4681 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg gcgtaatagc
     4741 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4801 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4861 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4921 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4981 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     5041 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     5101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5221 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     5281 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6061 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6121 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6181 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6241 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6301 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6361 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttc
//
