LOCUS       sequence-443051-        5898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated gene expression from CBh promoter. Contains canonical 
            left and right AAV2 ITRs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5898)
  TITLE     AAV ITR constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5898)
  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            "Unpublished"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..5898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   1016..1156
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1027..1156
                     /label=AAV2 ITR
     enhancer        1254..1557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1559..1837
                     /label=chicken beta-actin promoter
     intron          1838..2066
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     misc_feature    2841..3393
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2985..3002)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3212..3231
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3513..4101
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3566..3586)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3984..3995)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4128..4249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4139..4158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4193..4212
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4287..4427
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4287..4416
                     /label=AAV2 ITR
     primer_bind     complement(4498..4517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4617..4639
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4677..4695)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4763..4867
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4868..4936,4937..5728)
                     /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(5086..5105)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg catgccctgc
     1021 aggcagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
     1081 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     1141 atcactaggg gttcctgcgg ccgcagccgt gttttagagc tagaaatagc aagttaaaat
     1201 aaggctagtc cgtagcgcgt gcgccaattc tgcagacaaa ttctagaggt acccgttaca
     1261 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     1321 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     1381 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     1441 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     1501 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     1561 gaggtgagcc ccacgttctg cttcactctc cccatctccc cccccctccc cacccccaat
     1621 tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg
     1681 gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc
     1741 ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg
     1801 gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcga cgctgccttc
     1861 gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt
     1921 tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagctgagca
     1981 agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat tacctggagc
     2041 acctgcctga aatcactttt tttcaggttg gaccggtgcc gtttaaacaa caattggaag
     2101 agctcgtcga cccatggtgt ccgagctgat caaggagaac atgcacatga agctgtacat
     2161 ggagggcacc gtgaacaatc accacttcaa gtgcacatct gagggcgagg gcaagcccta
     2221 tgagggcacc cagacaatga ggatcaaggc agtggaggga ggaccactgc ctttcgcctt
     2281 tgacatcctg gccaccagct tcatgtacgg ctccaagacc tttatcaacc acacacaggg
     2341 catccccgac ttctttaagc agagcttccc tgagggcttt acctgggagc gcgtgaccac
     2401 atacgaggat ggcggcgtgc tgaccgccac acaggacaca tccctgcagg atggctgtct
     2461 gatctataat gtgaagatca ggggcgtgaa ctttccatct aatggccccg tgatgcagaa
     2521 gaagaccctg ggatgggagg caagcaccga gacactgtac ccagcagacg gaggcctgga
     2581 gggaagggcc gatatggccc tgaagctggt gggaggagga cacctgatct gcaacctgaa
     2641 gaccacatat agatctaaga agcctgccaa gaatctgaag atgccaggcg tgtactatgt
     2701 ggacaggaga ctggagcgga tcaaggaggc cgataaggag acatacgtgg agcagcacga
     2761 ggtggcagtg gcacggtatt gtgatctgcc ctccaagctg ggccaccgct gagaattcga
     2821 ttacggcggc ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc
     2881 gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa
     2941 acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat
     3001 gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac
     3061 aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg
     3121 cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt
     3181 tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg
     3241 gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac
     3301 atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga
     3361 cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatgctg gccggcctac
     3421 ttaattaacc actagttctg gatccttaaa actcgaggcc tataacgcgt atagctagct
     3481 ttgcgatcgc aaaggcgcgc cagcttatcg ataatcaacc tctggattac aaaatttgtg
     3541 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3601 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3661 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3721 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3781 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3841 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3901 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     3961 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4021 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     4081 ccctttgggc cgcctccccg catcgatacc gagcgctgga gagatcttaa gatacattga
     4141 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4201 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttc tgattttgta
     4261 gttaacacgt gcgaccgagc ggccgcagga acccctagtg atggagttgg ccactccctc
     4321 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4381 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4441 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4501 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     4561 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     4621 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     4681 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     4741 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4801 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4861 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4921 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4981 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5041 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5101 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5161 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5221 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5281 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5341 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5401 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5461 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5521 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5581 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5641 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5701 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5761 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5821 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5881 aaagatcaaa ggatcttc
//
