LOCUS       sequence-222921-        3882 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3882)
  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 3882)
  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-mir30
FEATURES             Location/Qualifiers
     source          1..3882
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     80..99
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     ncRNA           152..245
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(468..487)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    533..588
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(587..606)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(949..965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(949..965)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(962..984)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    973..989
                     /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(997..1003,1004..1021,1022..1027))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1042..1063
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1122..1317
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1168..1303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1323..1457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1360..1379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1414..1433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1525..1542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1696..2284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1776..1795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2455..3246,3247..3315))
                     /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     3078..3097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3316..3420)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3451..3830
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
       61 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
      121 gaatttcgac tagggataac agggtaattg tttgaatgag gcttcagtac tttacagaat
      181 cgttgcctgc acatcttgga aacacttgct gggattactt cttcaggtta acccaacaga
      241 aggctcgagc aaccagaatt caaggggcta ctttaggagc aattatcttg tttactaaaa
      301 ctgaatacct tgctatctct ttgatacatt tttacaaagc tgaattaaaa tggtataaat
      361 tatcactttt gatatcatcg atgcggccgc actcctcagg tgcaggctgc ctatcagaag
      421 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
      481 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
      541 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
      601 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
      661 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
      721 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
      781 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
      841 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
      901 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
      961 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1021 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1081 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     1141 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1201 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     1261 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     1321 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1381 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1441 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1501 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3061 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3421 cacatttccc cgaaaagtgc cacctgggtc gacattgatt attgactagt tattaatagt
     3481 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     3541 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     3601 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     3661 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     3721 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     3781 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
     3841 gccccacgtt ctgcttcact ctccccatct cccccccctc cc
//
