LOCUS       sequence-203639-        5863 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV-mediated gene expression from CBh promoter. Contains canonical 
            left and right AAV2 ITRs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5863)
  TITLE     AAV ITR constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5863)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CBh-mKate2-IRES-MCS (WT-WT)
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          127..355
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     misc_feature    1130..1682
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1274..1291)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1501..1520
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    1802..2390
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1855..1875)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2273..2284)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2417..2538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2428..2447)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2482..2501
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2576..2716
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2576..2705
                     /label=AAV2 ITR
     primer_bind     complement(2787..2806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2906..2928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2966..2984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3052..3156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3157..3225,3226..4017)
                     /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(3375..3394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4188..4776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4677..4696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5064..5085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5100..5105,5106..5123,5124..5130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5138..5154
                     /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)."
     primer_bind     5143..5165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5162..5178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5162..5178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   5203..5343
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5214..5343
                     /label=AAV2 ITR
     enhancer        5441..5744
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
       61 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      121 cgggcgggag tcgctgcgac gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      181 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      241 ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg
      301 tggggtatta atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg
      361 accggtgccg tttaaacaac aattggaaga gctcgtcgac ccatggtgtc cgagctgatc
      421 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tgaacaatca ccacttcaag
      481 tgcacatctg agggcgaggg caagccctat gagggcaccc agacaatgag gatcaaggca
      541 gtggagggag gaccactgcc tttcgccttt gacatcctgg ccaccagctt catgtacggc
      601 tccaagacct ttatcaacca cacacagggc atccccgact tctttaagca gagcttccct
      661 gagggcttta cctgggagcg cgtgaccaca tacgaggatg gcggcgtgct gaccgccaca
      721 caggacacat ccctgcagga tggctgtctg atctataatg tgaagatcag gggcgtgaac
      781 tttccatcta atggccccgt gatgcagaag aagaccctgg gatgggaggc aagcaccgag
      841 acactgtacc cagcagacgg aggcctggag ggaagggccg atatggccct gaagctggtg
      901 ggaggaggac acctgatctg caacctgaag accacatata gatctaagaa gcctgccaag
      961 aatctgaaga tgccaggcgt gtactatgtg gacaggagac tggagcggat caaggaggcc
     1021 gataaggaga catacgtgga gcagcacgag gtggcagtgg cacggtattg tgatctgccc
     1081 tccaagctgg gccaccgctg agaattcgat tacggcggcc ccccccctaa cgttactggc
     1141 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1201 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1261 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1321 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1381 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1441 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1501 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1561 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     1621 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
     1681 tatggccaca accatgctgg ccggcctact taattaacca ctagttctgg atccttaaaa
     1741 ctcgaggcct ataacgcgta tagctagctt tgcgatcgca aaggcgcgcc agcttatcga
     1801 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     1861 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     1921 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     1981 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2041 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2101 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2161 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     2221 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2281 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2341 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     2401 agcgctggag agatcttaag atacattgat gagtttggac aaaccacaac tagaatgcag
     2461 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2521 agctgcaata aacaagttct gattttgtag ttaacacgtg cgaccgagcg gccgcaggaa
     2581 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     2641 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     2701 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2761 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2821 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2881 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2941 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3001 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     4981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5041 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5161 acaggaaaca gctatgacca tgattacgcc aagcttgcat gccctgcagg cagctgcgcg
     5221 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     5281 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     5341 cctgcggccg cagccgtgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     5401 agcgcgtgcg ccaattctgc agacaaattc tagaggtacc cgttacataa cttacggtaa
     5461 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     5521 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     5581 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     5641 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     5701 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     5761 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     5821 ttttttaatt attttgtgca gcgatggggg cggggggggg ggg
//
