LOCUS       sequence-62258-e        7156 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 7156)
  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 7156)
  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-sGFP1-10::NLG1
FEATURES             Location/Qualifiers
     source          1..7156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     sig_peptide     621..680
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             684..710
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1841..1850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3944..4065
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3981..4000)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4035..4054
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4072..4527)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4230)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4421..4440
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4554..4658
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4660..5017
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4660..4851
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4683..4703)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4868..5003
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5052..5846
                     /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             5052..5846
                     /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(5106..5125)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5716..5735
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6034..6053)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6078..6125
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6454..7042
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6943..6962
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgaat
      601 tcggcttggg gatatccacc atggagacag acacactcct gctatgggta ctgctgctct
      661 gggttccagg ttccactggt gactatccat atgatgttcc agattatgct atgtccaaag
      721 gagaagagct gttcactgga gtggtaccaa tacttgtgga gttggacgga gatgtgaacg
      781 gacacaaatt ttcagtccgc ggggaggggg aaggggatgc tactattggc aagctgacgc
      841 tcaaattcat ctgtaccacc ggaaaactcc ctgtaccctg gcccacactg gtgacaactc
      901 tgacttacgg cgtgcaatgt tttagccgat acccagacca catgaagagg cacgactttt
      961 tcaaaagcgc aatgcctgaa ggatacgtac aggaaaggac catttctttt aaagacgacg
     1021 ggaagtacaa aacccgggca gtggtgaagt ttgagggcga taccctcgtc aataggatcg
     1081 aattgaaggg aactgacttc aaagaagatg gcaacatcct gggtcacaag cttgagtata
     1141 actttaactc ccacaacgtg tatattacag ccgacaaaca gaagaatgga attaaggcta
     1201 acttcactgt cagacacaat gtcgaagatg gctccgtgca gctcgccgat cactatcaac
     1261 agaatactcc tatcggggac ggcccagtcc tgctgcccga caaccactac ctgagtaccc
     1321 agactgttct gagcaaagat ccgaacgaga agggaaccgt cgaggggtct acatctggaa
     1381 gatctttgga tgatgtagac ccattggtta ctactaactt tggcaagatt aggggaatta
     1441 agaaagaact caataatgaa attttgggtc ctgtcattca gtttcttggg gttccatatg
     1501 ccgctccacc aacaggagaa catcgtttcc agcctccaga accaccatct ccctggtctg
     1561 acatccggaa cgccactcag tttgctcctg tatgtcccca gaatatcatt gatggcagat
     1621 tgcctgaagt tatgcttcct gtgtggttca ctaataactt ggatgtggtt tcatcatacg
     1681 tccaagacca gagtgaagac tgtctatact taaacatcta tgtcccaact gaagatggtc
     1741 cccttacaaa gaaacacaca gatgatttag gtgataatga cggtgctgaa gatgaagata
     1801 ttcgggacag tgggggtccc aaaccagtga tggtgtacat ccatggcggc tcttacatgg
     1861 aaggtactgg aaatctgtat gatgggagtg tcttggcaag ctatggcaat gtgatcgtca
     1921 tcacagtcaa ctatcggctt ggggtacttg gcttcttgag cacaggggat caggctgcca
     1981 aaggaaacta cgggctcctt gacctcatcc aggccctaag atggaccagc gagaacattg
     2041 ggttctttgg tggtgacccc ttgcgaatca ctgtgtttgg atcaggcgct gggggttcat
     2101 gtgtcaacct gctgacttta tcccattatt ctgaaggtaa ccgttggagc aattcaacca
     2161 aaggactttt tcaacgagca atagctcaga gtggaacagc cctttccagc tgggctgtta
     2221 gtttccagcc tgcaaaatac gctagaattc tggccacaaa agttggctgc aatgtttcag
     2281 atacagtaga gttagtagaa tgcctgcaga agaagcctta caaagaactt gttgatcaag
     2341 atgttcaacc agcccgatac cacatagcct ttggacctgt gatcgatggt gatgtaatac
     2401 cagatgaccc tcagatactg atggaacaag gagagttcct caactatgat ataatgttgg
     2461 gagttaacca aggggaaggg ttgaagtttg tcgaaaatat agtagatagt gatgatggtg
     2521 tatcagccag tgatttcgac tttgctgttt ctaattttgt tgataattta tatggatatc
     2581 ctgaaggcaa agatgttttg agagaaacca ttaaattcat gtatactgac tgggctgatc
     2641 gccataatcc tgaaactaga aggaagacat tgttggcttt gtttacggac catcaatggg
     2701 tagcacctgc tgtggccaca gcagaccttc actcgaactt tggctcacct acatacttct
     2761 atgcctttta tcatcattgc caaacagatc aagttccagc ttgggctgat gcagctcatg
     2821 gggatgaggt tccctatgtg ttgggaatcc ccatgattgg ccctacagag ttatttcctt
     2881 gcaatttctc caagaatgat gtgatgttga gtgcagtagt aatgacatac tggacgaatt
     2941 ttgctaaaac tggtgaccca aatcaaccag ttcctcaaga cacaaaattc atccatacca
     3001 aacccaaccg ctttgaagaa gtagcatgga ccagatattc ccagaaagac cagctttatc
     3061 tccatattgg attaaaaccg agagttaaag agcattacag agccaataag gtaaatctct
     3121 ggctggagct ggtacctcat ctgcataatc tcaatgacat ttctcagtat acctcgacaa
     3181 caactaaagt gccatccacg gacatcactc tcagacctac aaggaagaat tccacaccag
     3241 tcacatcagc ctttcccact gccaaacagg atgatcccaa gcaacaacca agccccttct
     3301 cggtggatca gagggactac tccacagagc taagtgtcac tatcgcagtg ggggcctctc
     3361 tgctgtttct caacatcttg gcttttgcag ccctgtacta caagaaggat aagaggagac
     3421 atgatgtcca ccggaggtgc agccctcagc gcacgaccac caacgaccta acccatgctc
     3481 cagaagagga aattatgtct ctccaaatga agcacactga cttggatcac gagtgtgagt
     3541 ccatccatcc acatgaggtg gttcttcgga ccgcctgtcc cccagattat actctagcta
     3601 tgaggaggtc acctgatgat attccactaa tgacacctaa caccatcaca atgattccca
     3661 acactatacc agggattcag cccttacata cattcaacac atttactgga ggacagaata
     3721 atacactgcc ccatccccac ccacaccccc attcacattc aacaaccagg gtatagaagc
     3781 ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat
     3841 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3901 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3961 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4021 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     4081 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4141 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4201 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4261 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4321 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4381 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4441 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4501 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4561 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4621 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4681 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4741 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4801 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4861 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4921 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4981 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     5041 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5101 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5161 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5221 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5281 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5341 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5401 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5461 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5521 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5581 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5641 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5701 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5761 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5821 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5881 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5941 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     6001 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6061 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6121 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6181 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6241 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6301 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6361 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6421 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6481 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6541 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6601 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6661 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6721 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6781 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6841 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6901 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6961 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     7021 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7081 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7141 tattaccgcc atgcat
//
