LOCUS       sequence-324014-        6957 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 6957)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Controlled expression of transgenes introduced by in vivo 
            electroporation.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Jan 16;104(3):1027-32. doi: 
            10.1073/pnas.0610155104
  PUBMED    17209010
REFERENCE   2  (bases 1 to 6957)
  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: "10.1073/pnas"; 
            journalName: "Proc Natl Acad Sci U S A"; date: "2007-01-16- 16"; 
            volume: "104"; issue: "3"; pages: "1027-32"
COMMENT     Sequence Label: pCAG-mGFP
FEATURES             Location/Qualifiers
     source          1..6957
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          337..1353
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1277..1299
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1361..1380
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(1495..1497,1498..1500,1501..2214)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1540..1561)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1801..1820)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2148..2169
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2317..2336)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2382..2437
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2436..2455)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2798..2814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2798..2814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2811..2833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2822..2838
                     /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(2846..2852,2853..2870,2871..2876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2891..2912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2971..3166
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3017..3152
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3172..3306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3209..3228)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3263..3282
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3374..3391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3545..4133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3625..3644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5645..6433,6434..6502))
                     /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
                     LIKH"
     primer_bind     6265..6284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6503..6607)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(6638..6957,1..60)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
       61 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
      121 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
      181 ggsgcgccgg gggggggggg gggggggggg gggggggggg gggsgrggsg grgrggkgcg
      241 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
      301 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      361 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      421 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      481 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      541 cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt ggggagcgcc
      601 gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
      661 gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg
      721 ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg agcagggggt
      781 gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt tgctgagcac
      841 ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc cgtgccgggc
      901 ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggrgggc
      961 tcgggggrgg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc ggcgagccgc
     1021 agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt gtcccaaatc
     1081 tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg cggggcgaag
     1141 cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc gccgcgccgc
     1201 cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg ccttcggggg
     1261 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     1321 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg
     1381 ctgtctcatc attttggcaa agaattcctc gaggatgatc caccggtcgc caccatgctg
     1441 tgctgtatga gaagaaccaa acaggttgaa aagaatgatg aggaccaaaa gatcatggtg
     1501 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1561 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     1621 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1681 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1741 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     1801 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1861 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1921 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1981 aaggcgaact tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac
     2041 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     2101 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     2161 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc
     2221 ccggaattat cgcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     2281 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     2341 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     2401 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2461 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     2521 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     2581 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     2641 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     2701 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag
     2761 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2821 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2881 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2941 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     3001 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3061 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3121 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     3181 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3241 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3301 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatcc
     4321 tgatgaatcc cctaatgatt tttatcaaaa tcattaagtt aaggtagata cacatcttgt
     4381 catatgatca aatggtttcg ccaaaaatca ataatcagac aacaaaatgt gcgaactcga
     4441 tattttacac gactctcttt accaattctg ccccgaatta cacttaaaac gactcaacag
     4501 cttaacgttg gcttgccacg ccttacttga ctgtaaaact ctcactctta ccgaacttgg
     4561 ccgtaacctg ccaaccaaag cgagaacaaa acataacatc aaacgaatcg accgattgtt
     4621 aggtaatcgt cacctccaca aagagcgact cgctgtatac cgttggcatg ctagctttat
     4681 ctgttcgggc aatacgatgc ccattgtact tgttgactgg tctgatatcc gtgagcaaaa
     4741 acggcttatg gtattgcgag cttcagtcgc actacacggt cgttctgtta ctctttatga
     4801 gaaagcgttc ccgctttcag agcaatgttc aaagaaagct catgaccaat ttctagccga
     4861 ccttgcgagc attctaccga gtaacaccac accgctcatt gtcagtgatg ctggctttaa
     4921 agtgccatgg tataaatccg ttgagaagct gggttggtac tggttaagtc gagtaagagg
     4981 aaaagtacaa tatgcagacc taggagcgga aaactggaaa cctatcagca acttacatga
     5041 tatgtcatct agtcactcaa agactttagg ctataagagg ctgactaaaa gcaatccaat
     5101 ctcatgccaa attctattgt ataaatctcg ctctaaaggc cgaaaaaatc agcgctcgac
     5161 acggactcat tgtcaccacc cgtcacctaa aatctactca gcgtcggcaa aggagccatg
     5221 gattctagca actaacttac ctgttgaaat tcgaacaccc aaacaacttg ttaatatcta
     5281 ttcgaagcga atgcagattg aagaaacctt ccgagacttg aaaagtcctg cctacggact
     5341 aggcctacgc catagccgaa cgagcagctc agagcgtttt gatatcatgc tgctaatcgc
     5401 cctgatgctt caactaacat gttggcttgc gggcgttcat gctcagaaac aaggttggga
     5461 caagcacttc caggctaaca cagtcagaaa tcgaaacgta ctctcaacag ttcgcttagg
     5521 catggaagtt ttgcggcatt ctggctacac aataacaagg gaagacttac tcgtggctgc
     5581 aaccctacta gctcaaaatt tattcacaca tggttacgct ttggggaaat tatgagggga
     5641 tctctcagtg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5701 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5821 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6061 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6601 ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt gactagttat
     6661 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6721 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6781 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6841 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6901 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatg
//
