LOCUS       sequence-51307-e        8678 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 8678)
  AUTHORS   Kay JN, Chu MW, Sanes JR
  TITLE     MEGF10 and MEGF11 mediate homotypic interactions required for mosaic
            spacing of retinal neurons.
  JOURNAL   Nature. 2012 Mar 11;483(7390):465-9. doi: 10.1038/nature10877.
  PUBMED    22407321
REFERENCE   2  (bases 1 to 8678)
  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"; journalName: 
            "Nature"; date: "2012-03-11- 11"; volume: "483"; issue: "7390"; 
            pages: "465-9"
COMMENT     Sequence Label: Ubc-megf10-GFP
FEATURES             Location/Qualifiers
     source          1..8678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..1257
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1094..1113
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     protein_bind    1325..1349
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1356..1367)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4846..4870)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      4890..4899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4896..4898,4899..4901,4902..5615)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4896..4898,4899..4901,4902..5612)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4941..4962)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5202..5221)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5549..5570
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5702..5721)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5767..5822
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5821..5840)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6181..6197)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6181..6197)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6194..6216)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6205..6221
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6229..6235,6236..6253,6254..6259))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6274..6295
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6354..6549
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6400..6535
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6555..6689
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6592..6611)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6646..6665
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6757..6774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6928..7516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7008..7027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7687..8478,8479..8547))
                     /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     8310..8329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8548..8652)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtaacccgt gtcggctcca gatctggcct ccgcgccggg
       61 ttttggcgcc tcccgcgggc gcccccctcc tcacggcgag cgctgccacg tcagacgaag
      121 ggcgcagcga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
      181 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
      241 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
      301 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
      361 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
      421 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
      481 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
      541 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
      601 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
      661 gtggggggca tggtgggcgg caagaaccca aggtcttgag gccttcgcta atgcgggaaa
      721 gctcttattc gggtgagatg ggctggggca ccatctgggg accctgacgt gaagtttgtc
      781 actgactgga gaactcgggt ttgtcgtctg gttgcggggg cggcagttat gcggtgccgt
      841 tgggcagtgc acccgtacct ttgggagcgc gcgcctcgtc gtgtcgtgac gtcacccgtt
      901 ctgttggctt ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc
      961 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     1021 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     1081 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     1141 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     1201 tgttagacta gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacgaa
     1261 gcttgggctg caggtcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggccca
     1321 tcaaacaagt ttgtacaaaa aagcaggctc cgaattcgcc cttcgattgt tcttcacaga
     1381 aaatggcgat ttcttcaagt tcgtgcctgg gcctcatctg ctcactgctc tgtcactggg
     1441 tggggacagc atcctccctg aacctggaag accccaacgt atgcagccac tgggaaagct
     1501 actcggtgac tgtgcaggag tcgtatccac atcccttcga tcagatctac tacacaagct
     1561 gcaccgacat cctgaactgg tttaaatgca cacggcacag aatcagctac cggacagcct
     1621 accgccacgg ggagaaaacc atgtatagac gcaaatccca gtgttgccca ggattttatg
     1681 aaagccgaga catgtgtgtc cctcactgtg ctgataaatg tgtccatggt cgctgcattg
     1741 ctccaaacac ctgtcagtgt gagcctggct ggggtgggac caactgtagc agtgcttgtg
     1801 atggtgatca ctgggggcct cactgcagca gccgatgcca gtgcaaaaac agagctttgt
     1861 gtaaccccat caccggtgct tgccactgcg ctgcgggcta ccggggatgg cgctgcgagg
     1921 accgttgtga acagggcacg tacggtaacg actgtcacca aagatgccag tgtcagaatg
     1981 gagcgacctg tgaccacatc actggggaat gccgttgttc acctgggtac actggagcct
     2041 tctgtgagga tctttgtcct cctggcaaac atggtccaca ttgtgagcag aggtgtccct
     2101 gccaaaatgg gggtgtgtgc caccatgtca ctggagagtg ctcttgccct tctggttgga
     2161 tgggcacagt gtgtggtcag ccctgccctg agggtcgctt tggaaagaac tgttcccaag
     2221 aatgccagtg tcacaatgga ggaacgtgtg atgctgccac aggccagtgt cactgcagcc
     2281 caggatacac aggggaacgg tgtcaggacg aatgtcctgt tgggagctat ggagttcgct
     2341 gtgctgaggc ctgcaggtgt gtcaacggag gcaagtgtta ccacgtgagt ggtacatgcc
     2401 tgtgcgaagc aggcttttcg ggtgaacttt gcgaggcgcg cctgtgtccc gaggggcttt
     2461 acggcatcaa atgtgacaag cggtgcccct gccacctgga caacactcac agctgtcatc
     2521 ccatgtctgg agagtgtggc tgcaagccgg gttggtcggg actgtactgt aatgaaacat
     2581 gctcccctgg attctacggg gaggcttgcc aacagatctg cagctgccag aacggggcgg
     2641 actgcgacag tgtgactgga aggtgtgcct gcgctccagg attcaaaggg actgactgct
     2701 ctactccgtg tcctctggga cgctacggga taaattgttc ttctcgctgt ggctgtaaaa
     2761 atgatgctgt ctgttctcct gtggatggat catgtatctg taaggcaggc tggcacgggg
     2821 tggactgttc catccgctgc cccagtggca catggggctt tggctgtaac ctaacgtgtc
     2881 agtgcctcaa tggcggtgcc tgcaacacgc tggatgggac ctgcacctgt gcgcccggat
     2941 ggcgaggcgc gaagtgtgaa tttccctgcc aggatggcac ttatgggctg aactgtgccg
     3001 agcgctgtga ctgcagccat gcagatggct gtcaccccac tacaggccat tgccgctgcc
     3061 tccctggatg gtcaggtgtg cactgtgaca gtgtgtgcgc tgagggacgc tggggtccta
     3121 actgctcgct gccctgctac tgtaaaaatg gggcttcgtg ttctccggat gatggcatct
     3181 gtgagtgtgc acccggattc cgaggcacca cttgccagag aatctgctcc cccggttttt
     3241 atggacatcg ctgtagccag acctgcccgc agtgtgtgca cagcagtggg ccctgccacc
     3301 acatcacggg cctgtgtgac tgcttacctg gcttcaccgg tgccctgtgc aatgaagtgt
     3361 gtcccagtgg cagatttggg aaaaactgtg caggcgtttg tacttgcacc aacaatggca
     3421 cctgtaaccc catcgacaga tcctgccagt gttacccagg ctggattggc agtgactgct
     3481 cccagccctg tccacctgcg cactggggtc cgaactgcat ccacacctgc aactgccaca
     3541 acggagcctt ttgcagcgcc tatgatgggg aatgcaaatg cactcctggc tggacggggc
     3601 tctactgcac tcagagatgc cctctgggct tctatggtaa ggactgtgca ctgatatgcc
     3661 aatgtcaaaa cggagctgac tgcgaccata tctcggggca gtgtacctgc cgcacgggat
     3721 tcatgggacg gcactgtgaa cagaagtgcc ctgcgggaac atacggctat ggctgtcgcc
     3781 agatctgtga ctgtctgaac aactccacct gtgaccacat cactggcacg tgttactgta
     3841 gcccaggatg gaaaggggca cgatgtgacc aagctggggt tatcatcgtg ggcaatctga
     3901 acagcttaag ccggaccagc accgcccttc ctgccgattc ctatcagatc ggggccatcg
     3961 cgggcatcgt ggtcctcgtt cttgttgtgc tcttcctgct ggcgctgttc atcatctaca
     4021 gacacaagca gaagaggaag gaatcaagca tgccggccgt gacctacacc cccgccatga
     4081 gggtcatcaa tgcagactat accatcgcag aaaccctgcc tcacagcaat ggtggaaatg
     4141 ccaacagcca ctactttacc aatcccagtt atcacacact tagccagtgt gccacatccc
     4201 ctcatgtgaa caatagggac aggatgacca ttgcaaagtc aaaaaacaat cagctgtttg
     4261 tgaatcttaa aaatgtgaat ccagggaaga gagggacatt ggtggactgc actgggacat
     4321 tgccagctga ctggaagcaa ggaggctacc tcaatgagct tggtgctttc gggctggaca
     4381 gaagctacat gggaaagtcc ttaaaagatc tggggaagaa ctctgaatat aattcaagca
     4441 cttgctcctt aagcagctct gaaaacccat atgccaccat taaagacccg cctgcactcc
     4501 tgcctaaaag ctccgagtgc ggctacgtgg agatgaagtc gccggcgcgg agagactccc
     4561 catatgcaga gatcaacaac tcaactccag ccaacaggaa tgtctatgaa gtcgaaccta
     4621 cagtgagcgt tgtgcaagga gtattcagca acagcggtca cgtcacccaa gacccatatg
     4681 accttccaaa gaacagtcac atcccttgcc attatgacct gctgccagta agggacagtt
     4741 catcctcccc aaagagagag gatggtggtg gcagcaacag caccagcagc aacagcacca
     4801 gcagcagcag cagcagcagt gaatcacatc aaaagggcga attcgaccca gctttcttgt
     4861 acaaagtggt tcgatgggat ccaccggtcg ccaccatggt gagcaagggc gaggagctgt
     4921 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     4981 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     5041 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     5101 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     5161 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     5221 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     5281 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     5341 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     5401 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     5461 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     5521 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     5581 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcactcct caggtgcagg
     5641 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     5701 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     5761 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     5821 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     5881 agagtttggc aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt
     5941 catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact
     6001 tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa
     6061 ttttccttac atgttttact agccagattt ttcctcctct cctgactact cccagtcata
     6121 gctgtccctc ttctcttatg aagatccctc gacctgcagc ccaagcttgg cgtaatcatg
     6181 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     6241 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     6301 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tccgcatctc
     6361 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     6421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     6481 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     6541 ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     6601 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     6661 tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc aacgcgcggg
     6721 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6781 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6841 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     7141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     7381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7861 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8101 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8341 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8581 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8641 aggggttccg cgcacatttc cccgaaaagt gccacctg
//
