LOCUS       sequence-391336-        7469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses V5-tagged EGFR in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7469)
  AUTHORS   Roh H, Dorner BG, Ting AY
  TITLE     Cell-Type-Specific Intracellular Protein Delivery with Inactivated 
            Botulinum Neurotoxin.
  JOURNAL   J Am Chem Soc. 2023 May 10;145(18):10220-10226. doi: 
            10.1021/jacs.3c01145. Epub 2023 Apr 26.
  PUBMED    37098770
REFERENCE   2  (bases 1 to 7469)
  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: 
            "10.1021/jacs.3c01145"; journalName: "J Am Chem Soc"; date: 
            "2023-05-10- 10"; volume: "145"; issue: "18"; pages: "10220-10226"
COMMENT     Sequence Label: pAAV_CMV::V5-EGFR
FEATURES             Location/Qualifiers
     source          1..7469
                     /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"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        851..1154
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1155..1358
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1309..1329
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1403..1422
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1403..1421
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     1441..1500
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             1501..1542
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5140..5157)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5146..5353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5382..5522
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5382..5511
                     /label=AAV2 ITR
     rep_origin      5597..6052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5684..5703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5894..5915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6069..6088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6188..6210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6248..6266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6334..6438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6439..6507,6508..7299)
                     /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"
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 cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgttgtaca tagtaatcaa ttacggggtc attagttcat agcccatata
      841 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      901 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      961 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     1021 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     1081 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     1141 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     1201 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     1261 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     1321 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
     1381 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccacc
     1441 atgtacagga tgcaactcct gtcttgcatt gcactaagtc ttgcacttgt cacgaattcg
     1501 ggcaagccaa tccctaaccc cctgctgggc ctggacagca ccaccggtct ggaggaaaag
     1561 aaagtttgcc aaggcacgag taacaagctc acgcagttgg gcacttttga agatcatttt
     1621 ctcagcctcc agaggatgtt caataactgt gaggtggtcc ttgggaattt ggaaattacc
     1681 tatgtgcaga ggaattatga tctttccttc ttaaagacca tccaggaggt ggctggttat
     1741 gtcctcattg ccctcaacac agtggagcga attcctttgg aaaacctgca gatcatcaga
     1801 ggaaatatgt actacgaaaa ttcctatgcc ttagcagtct tatctaacta tgatgcaaat
     1861 aaaaccggac tgaaggagct gcccatgaga aatttacagg aaatcctgca tggcgccgtg
     1921 cggttcagca acaaccctgc cctgtgcaac gtggagagca tccagtggcg ggacatagtc
     1981 agcagtgact ttctcagcaa catgtcgatg gacttccaga accacctggg cagctgccaa
     2041 aagtgtgatc caagctgtcc caatgggagc tgctggggtg caggagagga gaactgccag
     2101 aaactgacca aaatcatctg tgcccagcag tgctccgggc gctgccgtgg caagtccccc
     2161 agtgactgct gccacaacca gtgtgctgca ggctgcacag gcccccggga gagcgactgc
     2221 ctggtctgcc gcaaattccg agacgaagcc acgtgcaagg acacctgccc cccactcatg
     2281 ctctacaacc ccaccacgta ccagatggat gtgaaccccg agggcaaata cagctttggt
     2341 gccacctgcg tgaagaagtg tccccgtaat tatgtggtga cagatcacgg ctcgtgcgtc
     2401 cgagcctgtg gggccgacag ctatgagatg gaggaagacg gcgtccgcaa gtgtaagaag
     2461 tgcgaagggc cttgccgcaa agtgtgtaac ggaataggta ttggtgaatt taaagactca
     2521 ctctccataa atgctacgaa tattaaacac ttcaaaaact gcacctccat cagtggcgat
     2581 ctccacatcc tgccggtggc atttaggggt gactccttca cacatactcc tcctctggat
     2641 ccacaggaac tggatattct gaaaaccgta aaggaaatca cagggttttt gctgattcag
     2701 gcttggcctg aaaacaggac ggacctccat gcctttgaga acctagaaat catacgcggc
     2761 aggaccaagc aacatggtca gttttctctt gcagtcgtca gcctgaacat aacatccttg
     2821 ggattacgct ccctcaagga gataagtgat ggagatgtga taatttcagg aaacaaaaat
     2881 ttgtgctatg caaatacaat aaactggaaa aaactgtttg ggacctccgg tcagaaaacc
     2941 aaaattataa gcaacagagg tgaaaacagc tgcaaggcca caggccaggt ctgccatgcc
     3001 ttgtgctccc ccgagggctg ctggggcccg gagcccaggg actgcgtctc ttgccggaat
     3061 gtcagccgag gcagggaatg cgtggacaag tgcaaccttc tggagggtga gccaagggag
     3121 tttgtggaga actctgagtg catacagtgc cacccagagt gcctgcctca ggccatgaac
     3181 atcacctgca caggacgggg accagacaac tgtatccagt gtgcccacta cattgacggc
     3241 ccccactgcg tcaagacctg cccggcagga gtcatgggag aaaacaacac cctggtctgg
     3301 aagtacgcag acgccggcca tgtgtgccac ctgtgccatc caaactgcac ctacggatgc
     3361 actgggccag gtcttgaagg ctgtccaacg aatgggccta agatcccgtc catcgccact
     3421 gggatggtgg gggccctcct cttgctgctg gtggtggccc tggggatcgg cctcttcatg
     3481 cgaaggcgcc acatcgttcg gaagcgcacg ctgcggaggc tgctgcagga gagggagctt
     3541 gtggagcctc ttacacccag tggagaagct cccaaccaag ctctcttgag gatcttgaag
     3601 gaaactgaat tcaaaaagat caaagtgctg ggctccggtg cgttcggcac ggtgtataag
     3661 ggactctgga tcccagaagg tgagaaagtt aaaattcccg tcgctatcaa ggaattaaga
     3721 gaagcaacat ctccgaaagc caacaaggaa atcctcgatg aagcctacgt gatggccagc
     3781 gtggacaacc cccacgtgtg ccgcctgctg ggcatctgcc tcacctccac cgtgcaactc
     3841 atcacgcagc tcatgccctt cggctgcctc ctggactatg tccgggaaca caaagacaat
     3901 attggctccc agtacctgct caactggtgt gtgcagatcg caaagggcat gaactacttg
     3961 gaggaccgtc gcttggtgca ccgcgacctg gcagccagga acgtactggt gaaaacaccg
     4021 cagcatgtca agatcacaga ttttgggctg gccaaactgc tgggtgcgga agagaaagaa
     4081 taccatgcag aaggaggcaa agtgcctatc aagtggatgg cattggaatc aattttacac
     4141 agaatctata cccaccagag tgatgtctgg agctacgggg tgaccgtttg ggagttgatg
     4201 acctttggat ccaagccata tgacggaatc cctgccagcg agatctcctc catcctggag
     4261 aaaggagaac gcctccctca gccacccata tgtaccatcg atgtctacat gatcatggtc
     4321 aagtgctgga tgatagacgc agatagtcgc ccaaagttcc gtgagttgat catcgaattc
     4381 tccaaaatgg cccgagaccc ccagcgctac cttgtcattc agggggatga aagaatgcat
     4441 ttgccaagtc ctacagactc caacttctac cgtgccctga tggatgaaga agacatggac
     4501 gacgtggtgg atgccgacga gtacctcatc ccacagcagg gcttcttcag cagcccctcc
     4561 acgtcacgga ctcccctcct gagctctctg agtgcaacca gcaacaattc caccgtggct
     4621 tgcattgata gaaatgggct gcaaagctgt cccatcaagg aagacagctt cttgcagcga
     4681 tacagctcag accccacagg cgccttgact gaggacagca tagacgacac cttcctccca
     4741 gtgcctgaat acataaacca gtccgttccc aaaaggcccg ctggctctgt gcagaatcct
     4801 gtctatcaca atcagcctct gaaccccgcg cccagcagag acccacacta ccaggacccc
     4861 cacagcactg cagtgggcaa ccccgagtat ctcaacactg tccagcccac ctgtgtcaac
     4921 agcacattcg acagccctgc ccactgggcc cagaaaggca gccaccaaat tagcctggac
     4981 aaccctgact accagcagga cttctttccc aaggaagcca agccaaatgg catctttaag
     5041 ggctccacag ctgaaaatgc agaataccta agggtcgcgc cacaaagcag tgaatttatt
     5101 ggagcatgag aattcctcga gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     5161 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5221 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5281 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5341 ggcatgctgg ggaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     5401 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     5461 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     5521 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     5581 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5641 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     5701 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5761 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5821 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5881 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     5941 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     6001 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     6061 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6121 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6181 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6241 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6301 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6361 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6421 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6481 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6541 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6601 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6661 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6721 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6781 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6841 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6901 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6961 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     7021 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     7081 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7141 gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7201 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7261 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7321 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7381 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7441 gaccccgtag aaaagatcaa aggatcttc
//
