LOCUS       sequence-229520-        3274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EFS driven Multi Cloning Site-3 without the AgeI restriction enzyme 
            site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3274)
  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 3274)
  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: pJEP307-pAAV-EFS(No AgeI)-MCS3-pA
FEATURES             Location/Qualifiers
     source          1..3274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..32)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   71..211
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   71..200
                     /label=AAV2 ITR
     rep_origin      286..741
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(373..392)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..604
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(758..777)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     877..899
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(937..955)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1023..1127
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1128..1196,1197..1988)
                     /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(1346..1365)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2159..2747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2648..2667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2809..2938
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        2978..3189
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
ORIGIN
        1 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactggacc
       61 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
      121 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
      181 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
      241 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
      301 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      361 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      421 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      481 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
      541 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      601 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
      661 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
      721 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
      781 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
      841 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
      901 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
      961 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     1021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     1081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     1141 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     1201 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     1261 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     1321 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     1381 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     1441 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     1501 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     1561 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     1621 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     1681 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     1741 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     1801 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     1861 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     1921 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     1981 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     2041 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2101 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2161 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2221 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2281 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2341 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2401 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2461 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2521 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2581 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2641 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     2701 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     2761 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
     2821 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
     2881 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
     2941 ggccgcacgc gttcgagtgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
     3001 gtccccgaga agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc
     3061 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
     3121 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
     3181 agaacacagg tgtcgtgacg cgggaattcg gtacctctag agcaccggtc atatggatat
     3241 ctgactagtc tcgagtttat tgcagcttat aatg
//
