LOCUS       sequence-229569-        3608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CMV driven Multi Cloning Site-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3608)
  AUTHORS   Kumar N, Stanford W, de Solis C, Aradhana, Abraham ND, Dao TJ, 
            Thaseen S, Sairavi A, Gonzalez CU, Ploski JE
  TITLE     The Development of an AAV-Based CRISPR SaCas9 Genome Editing System 
            That Can Be Delivered to Neurons in vivo and Regulated via 
            Doxycycline and Cre-Recombinase.
  JOURNAL   Front Mol Neurosci. 2018 Nov 13;11:413. doi: 
            10.3389/fnmol.2018.00413. eCollection 2018.
  PUBMED    30483052
REFERENCE   2  (bases 1 to 3608)
  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: "Front Mol 
            Neurosci."; date: "2018-11-13"; volume: "11:413. doi"; pages: " 
            10.3389/fnmol.2018.00413. eCollection 2018"
COMMENT     Sequence Label: pJEP300-pAAV-CMV-MCS2-pA
FEATURES             Location/Qualifiers
     source          1..3608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        154..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        458..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     658..682
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     complement(813..832)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   871..1011
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   871..1000
                     /label=AAV2 ITR
     rep_origin      1086..1541
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1173..1192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1383..1404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1558..1577)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1677..1699
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1737..1755)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1823..1927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1928..1996,1997..2788)
                     /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     complement(2146..2165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2959..3547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3448..3467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cagtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      721 aagacaccgg aattcggtac caccggtgct ctagacatat ggatatctga ctagtctcga
      781 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      841 agcatttttt tcactggacc gagcggccgc aggaacccct agtgatggag ttggccactc
      901 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
      961 gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga
     1021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc
     1081 atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1141 gaccgctaca cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct
     1201 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1261 atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag
     1321 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1381 tagtggactc ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga
     1441 tttataaggg attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa
     1501 atttaacgcg aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac
     1561 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     1621 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     1681 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     1741 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1801 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1861 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1921 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1981 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2041 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2101 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     2161 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2221 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     2281 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     2341 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     2401 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     2461 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     2521 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     2581 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     2641 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2701 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2761 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2821 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2881 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2941 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3001 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3061 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     3121 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3181 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3241 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3301 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3361 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3421 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3481 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3541 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3601 tcacatgt
//
