LOCUS       sequence-200621-        5897 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 5897)
  TITLE     AAV ITR constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5897)
  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            "Unpublished"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..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   245..385
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   256..385
                     /label=AAV2 ITR
     enhancer        483..786
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        788..1065
                     /label=chicken beta-actin promoter
     intron          1066..1294
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1342..2040
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    2069..2621
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2213..2230)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2440..2459
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    2741..3329
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2794..2814)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3212..3223)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3356..3477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3367..3386)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3421..3440
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3515..3655
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3515..3644
                     /label=AAV2 ITR
     primer_bind     complement(3726..3745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3845..3867
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3905..3923)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3991..4095
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4096..4164,4165..4956)
                     /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(4314..4333)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5127..5715
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5616..5635
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgccctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
      301 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      361 gccaactcca tcactagggg ttcctgcggc cgcagccgtg ttttagagct agaaatagca
      421 agttaaaata aggctagtcc gtagcgcgtg cgccaattct gcagacaaat tctagaggta
      481 cccgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      541 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      601 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      661 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      721 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      781 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      841 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      901 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
      961 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     1021 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgacg
     1081 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
     1141 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     1201 gctgagcaag aggtaagggt ttaagggatg gttggttggt ggggtattaa tgtttaatta
     1261 cctggagcac ctgcctgaaa tcactttttt tcaggttgga ccggtgccgt ttaaacaaca
     1321 attggaagag ctcgtcgacc catggtgagc gagctgatta aggagaacat gcacatgaag
     1381 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
     1441 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
     1501 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
     1561 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
     1621 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     1681 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     1741 atgcagaaga aaacactcgg ctgggaggcc agcaccgaga ccctgtaccc cgctgacggc
     1801 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1861 aacttgaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1921 tactatgtgg acagaagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1981 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagatga
     2041 gaattcgatt acggcggccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     2101 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     2161 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     2221 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     2281 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     2341 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     2401 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     2461 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     2521 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac
     2581 cacggggacg tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatgctggc
     2641 cggcctactt aattaaccac tagttctgga tccttaaaac tcgaggccta taacgcgtat
     2701 agctagcttt gcgatcgcaa aggcgcgcca gcttatcgat aatcaacctc tggattacaa
     2761 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     2821 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     2881 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     2941 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3001 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3061 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     3121 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     3181 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     3241 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     3301 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctggaga gatcttaaga
     3361 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3421 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttctg
     3481 attttgtagt taacacgtgc gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc
     3541 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     3601 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     3661 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     3721 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3781 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3841 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3901 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3961 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4021 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4081 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4141 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4201 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4261 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4321 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4381 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4441 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4501 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4561 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4621 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4681 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4741 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4801 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4861 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4921 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4981 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5041 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5101 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5161 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5221 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5281 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5341 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5401 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5461 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5521 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5581 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5641 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5701 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5761 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5821 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5881 agcgaggaag cggaaga
//
