LOCUS       sequence-62259-e        5295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5295)
  AUTHORS   Yamagata M, Sanes JR
  TITLE     Transgenic strategy for identifying synaptic connections in mice by 
            fluorescence complementation (GRASP).
  JOURNAL   Front Mol Neurosci. 2012;5:18. Epub 2012 Feb 16.
  PUBMED    22355283
REFERENCE   2  (bases 1 to 5295)
  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: "Front Mol 
            Neurosci."; date: "2012"; volume: "5"; pages: "18. Epub 2012 Feb 16"
COMMENT     Sequence Label: m-sGFP11::NXN
FEATURES             Location/Qualifiers
     source          1..5295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        285..488
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     439..459
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             728..775
                     /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"
     polyA_signal    2083..2204
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2120..2139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2174..2193
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2211..2666)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2348..2369)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2560..2579
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2693..2797
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2799..3156
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2799..2990
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2822..2842)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3007..3142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3191..3985
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3191..3985
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3245..3264)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3855..3874
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4173..4192)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4217..4264
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4593..5181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5082..5101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
       61 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      121 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      181 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      241 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      301 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      361 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      421 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      481 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag atctcgagat
      541 gctccggtgc ggcgccgatc tgggatcgcc cgggggcggc agtggcggcg gcgcaggggg
      601 gcgcctggcc ctgatctgga tagtcccgct caccctcggc ggcctcctag gagtggcctg
      661 gggggcatcc agtttgggcg cgcacaccgg tgacacatac agatacataa gcgaggttac
      721 agcctctcgc gaccacatgg tgctgcatga gtatgtcaac gctgcgggaa ttaccggggg
      781 tggtgggtct ggtggagggg gatcccacat ccaccatttc catggcagca gcaagcatca
      841 ttcagtgcct attgcaatct acaggtcacc agcatccttg cgaggcggac acgctgggac
      901 aacatatatc tttagcaaag gtggtggaca gattacatat aagtggcctc ccaatgaccg
      961 ccccagtaca cgagcagaca ggctggccat cggatttagc actgtccaga aggaagccgt
     1021 gttggtgcgt gtggacagtt cctcaggcct gggtgactac ctggagctgc acatacacca
     1081 aggaaaaatt ggagttaagt ttaatgttgg gacagatgac atcgccatcg aagagtctaa
     1141 tgcaatcatt aatgatggga aataccatgt agtacgtttc acaaggagtg gtggcaatgc
     1201 cacgttacag gtggacagct ggccagttat cgaacgctac cctgcagggc gtcagctcac
     1261 aatcttcaat agccaagcaa ccataataat tggcgggaaa gagcagggcc agcccttcca
     1321 gggccagctc tctggtcttt actacaatgg cttgaaagtt ctgaatatgg cagcagagaa
     1381 cgatgccaac atcgccatag tggggaatgt gaggctggtc ggtgaagtgc cttcctctat
     1441 gacaactgag tcgacagcca ctgccatgca gtccgagatg tccacctcaa tcatggagac
     1501 caccacaacc ctggctacca gcacagctcg acgaggaaag ccccccacaa aggagcctat
     1561 cagccagacc acggatgata tccttgtggc ctcggcagag tgtcccagcg acgatgagga
     1621 cattgacccc tgtgagccga gctcaggtgg gttagccaac cccacccgag tgggcggccg
     1681 cgaaccatac ccaggctcgg cagaggtgat ccgggagtct agcagtacca ctggcatggt
     1741 ggtggggatt gtcgcagcag cagctctgtg catcctcatc ctcctctatg ccatgtacaa
     1801 gtacagaaac cgggatgaag gctcatatca cgtggatgag agtcgaaact acatcagtaa
     1861 ctcagcacag tccaatgggg ctgtggtcaa ggagaagcag cccagcagtg cgaaaagcgc
     1921 caacaaaaac aagaagaaca aggataagga gtattatgtc tagggatccg actctagatc
     1981 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     2041 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     2101 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     2161 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag
     2221 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     2281 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     2341 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     2401 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     2461 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     2521 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     2581 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2641 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     2701 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2761 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     2821 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2881 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2941 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3001 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3061 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3121 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     3181 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     3241 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3301 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3361 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3421 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3481 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3541 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3601 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3661 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     3721 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3781 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3841 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3901 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3961 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4021 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4081 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4141 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     4201 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     4261 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     4321 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     4381 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     4441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5281 attaccgcca tgcat
//
