pYW475

LOCUS      pYW475               5648 bp DNA     circular SYN 05-JUN-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5648)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 17, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     246..265
                     /label=OPQD895
     enhancer        261..640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        336..640
                     /label=CMV enhancer
                     /note="human cytomegalovirus (CMV) immediate early 
                     enhancer"
     promoter        641..844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      883..892
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             889..2541
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     1444..1463
                     /label=FLUC-QPCR-F
     primer_bind     complement(1567..1586)
                     /label=FLUC-QPCR-R
     primer_bind     complement(2522..2542)
                     /label=OPQD896
     polyA_signal    2556..2780
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2798..2938
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2798..2927
                     /label=AAV2 ITR
     primer_bind     complement(2996..3012)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3020..3036
                     /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)."
     promoter        complement(3044..3074)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3089..3110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3398..3986)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4157..5017)
                     /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"
     promoter        complement(5018..5122)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5596..5612
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct cgacaattgc atgaaggcga tgtacgggcc agatatacgc gtaatctgct
      181 tagggcggcc gccatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      841 agctctctgg ctaactagag aacccagcta gcgctaccgg tcgccaccat ggaagatgcc
      901 aaaaacatta agaagggccc agcgccattc tacccactcg aagacgggac cgccggcgag
      961 cagctgcaca aagccatgaa gcgctacgcc ctggtgcccg gcaccatcgc ctttaccgac
     1021 gcacatatcg aggtggacat tacctacgcc gagtacttcg agatgagcgt tcggctggca
     1081 gaagctatga agcgctatgg gctgaataca aaccatcgga tcgtggtgtg cagcgagaat
     1141 agcttgcagt tcttcatgcc cgtgttgggt gccctgttca tcggtgtggc tgtggcccca
     1201 gctaacgaca tctacaacga gcgcgagctg ctgaacagca tgggcatcag ccagcccacc
     1261 gtcgtattcg tgagcaagaa agggctgcaa aagatcctca acgtgcaaaa gaagctaccg
     1321 atcatacaaa agatcatcat catggatagc aagaccgact accagggctt ccaaagcatg
     1381 tacaccttcg tgacttccca tttgccaccc ggcttcaacg agtacgactt cgtgcccgag
     1441 agcttcgacc gggacaaaac catcgccctg atcatgaaca gtagtggcag taccggattg
     1501 cccaagggcg tagccctacc gcaccgcacc gcttgtgtcc gattcagtca tgcccgcgac
     1561 cccatcttcg gcaaccagat catccccgac accgctatcc tcagcgtggt gccatttcac
     1621 cacggcttcg gcatgttcac cacgctgggc tacttgatct gcggctttcg ggtcgtgctc
     1681 atgtaccgct tcgaggagga gctattcttg cgcagcttgc aagactataa gattcaatct
     1741 gccctgctgg tgcccacact atttagcttc ttcgctaaga gcactctcat cgacaagtac
     1801 gacctaagca acttgcacga gatcgccagc ggcggggcgc cgctcagcaa ggaggtaggt
     1861 gaggccgtgg ccaaacgctt ccacctacca ggcatccgcc agggctacgg cctgacagaa
     1921 acaaccagcg ccattctgat cacccccgaa ggggacgaca agcctggcgc agtaggcaag
     1981 gtggtgccct tcttcgaggc taaggtggtg gacttggaca ccggtaagac actgggtgtg
     2041 aaccagcgcg gcgagctgtg cgtccgtggc cccatgatca tgagcggcta cgttaacaac
     2101 cccgaggcta caaacgctct catcgacaag gacggctggc tgcacagcgg cgacatcgcc
     2161 tactgggacg aggacgagca cttcttcatc gtggaccggc tgaagagcct gatcaaatac
     2221 aagggctacc aggtagcccc agccgaactg gagagcatcc tgctgcaaca ccccaacatc
     2281 ttcgacgccg gggtcgccgg cctgcccgac gacgatgccg gcgagctgcc cgccgcagtc
     2341 gtcgtgctgg aacacggtaa aaccatgacc gagaaggaga tcgtggacta tgtggccagc
     2401 caggttacaa ccgccaagaa gctgcgcggt ggtgttgtgt tcgtggacga ggtgcctaaa
     2461 ggactgaccg gcaagttgga cgcccgcaag atccgcgaga ttctcattaa ggccaagaag
     2521 ggcggcaaga tcgccgtgta agaattctgc ctcgactgtg ccttctagtt gccagccatc
     2581 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2641 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2701 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2761 ggatgcggtg ggctctatgg ggcctcgagc gcgtgtgagg aacccctagt gatggagttg
     2821 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     2881 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcaggat
     2941 cggatgccgg gaccgacgag tgcagaggcg tgcaagcgag cttggcgtaa tcatggtcat
     3001 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3061 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3121 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3181 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3241 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3301 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3361 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3421 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3481 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3541 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3601 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3661 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3721 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3781 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3841 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3901 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3961 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4021 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4081 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4141 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4201 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4261 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4321 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4381 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4441 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4501 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4561 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4621 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4681 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4741 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4801 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4861 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4921 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4981 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5041 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5101 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     5161 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     5221 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5281 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5341 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     5401 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     5461 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     5521 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     5581 cccagtcacg acgttgtaaa acgacggcca gtgaattgga gatcggtact tcgcgaatgc
     5641 gtcgagat
//