LOCUS       sequence-350211-        8153 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 8153)
  AUTHORS   Klessner JL, Desai BV, Amargo EV, Getsios S, Green KJ
  TITLE     EGFR and ADAMs cooperate to regulate shedding and endocytic 
            trafficking of the desmosomal cadherin desmoglein 2.
  JOURNAL   Mol Biol Cell. 2009 Jan;20(1):328-37. Epub 2008 Nov 5.
  PUBMED    18987342
REFERENCE   2  (bases 1 to 8153)
  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: "Mol Biol 
            Cell."; date: "2009-01"; volume: "20(1)"; pages: "328-37. Epub 2008 
            Nov 5"
COMMENT     Sequence Label: 1105-Desmoglein2-FLAG-eGFP
FEATURES             Location/Qualifiers
     source          1..8153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        2..305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     460..480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    532..612
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(602..618)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(602..618)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3992..4015
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      4034..4043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4040..4042,4043..4045,4046..4759)
                     /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(4040..4042,4043..4045,4046..4756)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4085..4106)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4346..4365)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4693..4714
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4882..5003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4919..4938)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4973..4992
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5010..5465)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5168)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5359..5378
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5492..5596
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5598..5955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5598..5789
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5621..5641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5806..5941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5990..6784
                     /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             5990..6784
                     /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(6044..6063)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6654..6673
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6972..6991)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7016..7063
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7392..7980
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7881..7900
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
       61 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      121 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      181 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      241 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      301 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      361 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      421 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      481 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
      541 accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac cgcgggcccg
      601 ggatccacta gttctagagc ggccgctcga gggtgcgatg gcgcggagcc cgggacgcgc
      661 gtacgccctg ctgcttctcc tgatctgctt taacgttgga agtggacttc acttacaggt
      721 cttaagcaca agaaatgaaa ataagctgct tcctaaacat cctcatttag tgcggcaaaa
      781 gcgcgcctgg atcaccgccc ccgtggctct tcgggaggga gaggatctgt ccaagaagaa
      841 tccaattgcc aagatacatt ctgatcttgc agaagaaaga ggactcaaaa ttacttacaa
      901 atacactgga aaagggatta cagagccacc ttttggtata tttgtcttta acaaagatac
      961 tggagaactg aatgttacca gcattcttga tcgagaagaa acaccatttt ttctgctaac
     1021 aggttacgct ttggatgcaa gaggaaacaa tgtagagaaa cccttagagc tacgcattaa
     1081 ggttcttgat atcaatgaca acgaaccagt gttcacacag gatgtctttg ttgggtctgt
     1141 tgaagagttg agtgcagcac atactcttgt gatgaaaatc aatgcaacag atgcagatga
     1201 gcccaatacc ctgaattcga aaatttccta tagaatcgta tctctggagc ctgcttatcc
     1261 tccagtgttc tacctaaata aagatacagg agagatttat acaaccagtg ttaccttgga
     1321 cagagaggaa cacagcagct acactttgac agtagaagca agagatggca atggagaagt
     1381 tacagacaaa cctgtaaaac aagctcaagt tcagattcgt attttggatg tcaatgacaa
     1441 tatacctgta gtagaaaata aagtgcttga agggatggtt gaagaaaatc aagtcaatgt
     1501 agaagttacg cgcataaaag tgttcgatgc agatgaaata ggttctgata attggctggc
     1561 aaattttaca tttgcatcag gaaatgaagg aggttatttc cacatagaaa cagatgctca
     1621 aactaacgaa ggaattgtga cccttattaa ggaagtagat tatgaagaaa tgaagaatct
     1681 tgacttcagt gttattgtcg ctaataaagc agcttttcac aagtcgatta ggagtaaata
     1741 caagcctaca cccattccca tcaaggtcaa agtgaaaaat gtgaaagaag gcattcattt
     1801 taaaagcagc gtcatctcaa tttatgttag cgagagcatg gatagatcaa gcaaaggcca
     1861 aataattgga aattttcaag cttttgatga ggacactgga ctaccagccc atgcaagata
     1921 tgtaaaatta gaagatagag ataattggat ctctgtggat tctgtcacat ctgaaattaa
     1981 acttgcaaaa cttcctgatt ttgaatctag atatgttcaa aatggcacat acactgtaaa
     2041 gattgtggcc atatcagaag attatcctag aaaaaccatc actggcacag tccttatcaa
     2101 tgttgaagac atcaacgaca actgtcccac actgatagag cctgtgcaga caatctgtca
     2161 cgatgcagag tatgtgaatg ttactgcaga ggacctggat ggacacccaa acagtggccc
     2221 tttcagtttc tccgtcattg acaaaccacc tggcatggca gaaaaatgga aaatagcacg
     2281 ccaagaaagt accagtgtgc tgctgcaaca aagtgagaaa aagcttggga gaagtgaaat
     2341 tcagttcctg atttcagaca atcagggttt tagttgtcct gaaaagcagg tccttacact
     2401 cacagtttgt gagtgtctgc atggcagcgg ctgcagggaa gcacagcatg actcctatgt
     2461 gggcctggga cccgcagcaa ttgcgctcat gattttggcc tttctgctcc tgctattggt
     2521 accactttta ctgctgatgt gccattgcgg aaagggcgcc aaaggcttta cccccatacc
     2581 tggcaccata gagatgctgc atccttggaa taatgaagga gcaccacctg aagacaaggt
     2641 ggtgccatca tttctgccag tggatcaagg gggcagtcta gtaggaagaa atggagtagg
     2701 aggtatggcc aaggaagcca cgatgaaagg aagtagctct gcttccattg tcaaagggca
     2761 acatgagatg tccgagatgg atggaaggtg ggaagaacac agaagcctgc tttctggtag
     2821 agctacccag tttacagggg ccacaggcgc tatcatgacc actgaaacca cgaagaccgc
     2881 aagggccaca ggggcttcca gagacatggc cggagctcag gcagctgctg ttgcactgaa
     2941 cgaagaattc ttaagaaatt atttcactga taaagcggcc tcttacactg aggaagatga
     3001 aaatcacaca gccaaagatt gccttctggt ttattctcag gaagaaactg aatcgctgaa
     3061 tgcttctatt ggttgttgca gttttattga aggagagcta gatgaccgct tcttagatga
     3121 tttgggactt aaattcaaga cgctagctga agtttgcctg ggtcaaaaaa tagatataaa
     3181 taaggaaatt gagcagagac aaaaacctgc cacagaaaca agtatgaaca cagcttcaca
     3241 ttcactctgt gagcaaacta tggttaattc agagaatacc tactcctctg gcagtagctt
     3301 cccagttcca aaatctttgc aagaagccaa tgcagagaaa gtaactcagg aaatagtcac
     3361 tgaaagatct gtgtcttcta ggcaggcgca aaaggtagct acacctcttc ctgacccaat
     3421 ggcttctaga aatgtgatag caacagaaac ttcctatgtc acagggtcca ctatgccacc
     3481 aaccactgtg atcctgggtc ctagccagcc acagagcctt attgtgacag agagggtgta
     3541 tgctccagct tctaccttgg tagatcagcc ttatgctaat gaaggtacag ttgtggtcac
     3601 tgaaagagta atacagcctc atgggggtgg atcgaatcct ctggaaggca ctcagcatct
     3661 tcaagatgta ccttacgtca tggtgaggga aagagagagc ttccttgccc ccagctcagg
     3721 tgtgcagcct actctggcca tgcctaatat agcagtagga cagaatgtga cagtgacaga
     3781 aagagttcta gcacctgctt ccactctgca atccagttac cagattccca ctgaaaattc
     3841 tatgacggct aggaacacca cggtgtctgg agctggagtc cctggccctc tgccagattt
     3901 tggtttagag gaatctggtc attctaattc taccataacc acatcttcca ccagagtcac
     3961 caagcatagc actgtacagc attcttactc cgattacaag gacgacgatg acaaggggat
     4021 ccaccggccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4081 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4141 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4201 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4261 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4321 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4381 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4441 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     4501 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     4561 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4621 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     4681 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4741 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     4801 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     4861 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     4921 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4981 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     5041 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5101 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5161 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5221 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5281 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5341 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     5401 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     5461 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     5521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5581 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     5641 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5701 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5761 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5941 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     6001 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     6061 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     6121 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     6181 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     6241 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     6301 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     6361 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     6421 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     6481 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     6541 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     6601 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     6661 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     6721 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     6781 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     6841 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     6901 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     6961 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     7021 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     7081 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     7141 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     7201 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     7261 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7321 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7381 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7441 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7501 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     7561 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7621 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7681 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7741 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7801 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7861 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7921 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7981 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     8041 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
     8101 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tcc
//
