pYW470

LOCUS       pYW470              5675 bp DNA     circular SYN 03-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Macapp
  TITLE     Direct Submission
  JOURNAL   Exported Jun 22, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     complement(32..79)
                     /label=oYW787
                     /note="AAV ITR PCR primer2"
     primer_bind     complement(65..123)
                     /label=oYW788
                     /note="AAV ITR PCR primer3"
     primer_bind     135..157
                     /label=oYW1060
                     /note="pYW469 seq primer"
     primer_bind     172..192
                     /label=QL 408-F
     promoter        213..893
                     /label=PhGfaABC1D
     primer_bind     complement(395..414)
                     /label=oYW15
                     /note="pYW06-pGFAP (for Colony PCR) R"
     primer_bind     838..854
                     /label=oYW266
                     /note="PhGfaABC1D seq primer"
     primer_bind     856..875
                     /label=oYW1008
                     /note="PhGfaABC1D seq primer"
     primer_bind     complement(878..919)
                     /label=oYW279
                     /note="RA-NheI-P_GFAP-R"
     primer_bind     883..944
                     /label=oYW294
                     /note="LA-AgeI-F"
     primer_bind     895..922
                     /label=oYW1100
                     /note="LA-XbaI-PCR-F"
     regulatory      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             916..2568
                     /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     complement(945..962)
                     /label=GLP4
     primer_bind     1087..1105
                     /label=oYW232
                     /note="fLuc-qPCR-F"
     primer_bind     complement(1300..1320)
                     /label=oYW233
                     /note="fLuc-qPCR-R"
     primer_bind     1536..1553
                     /label=oYW363
                     /note="fLuc mid seq primer"
     primer_bind     complement(1698..1716)
                     /label=oYW216
                     /note="fLuciferase sequencing primer"
     primer_bind     1763..1780
                     /label=oYW442
                     /note="fLuc mid seq primer"
     primer_bind     complement(1793..1815)
                     /label=oYW1088
                     /note="fLuc seq primer"
     primer_bind     complement(2266..2275)
                     /label=oYW232
                     /note="fLuc-qPCR-F"
     primer_bind     2297..2315
                     /label=oYW473
                     /note="fLuc seq primer"
     primer_bind     complement(2577..2594)
                     /label=BGH/pcDNA3.1R
     primer_bind     complement(2577..2594)
                     /label=pcDNA3.1-R(BGH-R)
     primer_bind     2580..2599
                     /label=oYW188
                     /note="bGH poly A"
     polyA_signal    2583..2807
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2587..2604)
                     /label=oYW190
                     /note="BGH poly A R"
     primer_bind     complement(2649..2667)
                     /label=oYW57
                     /note="bGH seq primer R"
     primer_bind     complement(2652..2675)
                     /label=oYW302
                     /note="bGH pA R"
     primer_bind     2652..2671
                     /label=W2F0627-P099-D03
                     /note="Sequencing primer"
     primer_bind     2710..2729
                     /label=BGH-polyA-R
     repeat_region   2825..2965
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2825..2954
                     /label=AAV2 ITR
     primer_bind     2832..2890
                     /label=oYW788
                     /note="AAV ITR PCR primer3"
     primer_bind     2876..2934
                     /label=oYW787
                     /note="AAV ITR PCR primer2"
     primer_bind     complement(3022..3039)
                     /label=M13R
     primer_bind     complement(3023..3039)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3035..3058)
                     /label=M13R-48
     primer_bind     complement(3037..3060)
                     /label=pQE65
     primer_bind     complement(3041..3060)
                     /label=pQE30-F
     protein_bind    3047..3063
                     /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(3071..3101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3088..3107)
                     /label=PCAMBIA1301-F
     protein_bind    3116..3137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3380..3399)
                     /label=oYW207
                     /note="pLL3.7 PCR R"
     primer_bind     complement(3410..3423)
                     /label=oYW49
                     /note="ColE1 origin R for PBS-C/D box elongation"
     rep_origin      complement(3425..4013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3564..3583)
                     /label=W5CF0703-P099-D09
                     /note="Sequencing primer"
     CDS             complement(4184..5044)
                     /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(4201..4220)
                     /label=W4CF0701-P099-E08
                     /note="Sequencing primer"
     primer_bind     complement(4865..4884)
                     /label=W3CF0630-P099-D08
                     /note="Sequencing primer"
     primer_bind     complement(4921..4940)
                     /label=W3CF0630-P099-A08
                     /note="Sequencing primer"
     primer_bind     complement(4921..4940)
                     /label=W3CF0630-P099-F07
                     /note="Sequencing primer"
     primer_bind     5015..5034
                     /label=oYW24
                     /note="Forward seq primer of AmpR"
     promoter        complement(5045..5149)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5164..5183
                     /label=PBR322F
     primer_bind     5225..5244
                     /label=oYW18
                     /note="TET07 PCR F"
     primer_bind     complement(5273..5295)
                     /label=pGEX-3
     primer_bind     5572..5591
                     /label=P3/Pmal-C2X-R
     primer_bind     5579..5598
                     /label=PCAMBIA1301-R
     primer_bind     5596..5619
                     /label=M13F-47
     primer_bind     5622..5639
                     /label=M13F
     primer_bind     5623..5639
                     /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 gctacgggcc agatatacgc gtgaacatat cctggtgtgg agtaggggac
      241 gctgctctga cagaggctcg ggggcctgag ctggctctgt gagctgggga ggaggcagac
      301 agccaggcct tgtctgcaag cagacctggc agcattgggc tggccgcccc ccagggcctc
      361 ctcttcatgc ccagtgaatg actcaccttg gcacagacac aatgttcggg gtgggcacag
      421 tgcctgcttc ccgccgcacc ccagcccccc tcaaatgcct tccgagaagc ccattgagca
      481 gggggcttgc attgcacccc agcctgacag cctggcatct tgggataaaa gcagcacagc
      541 cccctagggg ctgcccttgc tgtgtggcgc caccggcggt ggagaacaag gctctattca
      601 gcctgtgccc aggaaagggg atcaggggat gcccaggcat ggacagtggg tggcaggggg
      661 ggagaggagg gctgtctgct tcccagaagt ccaaggacac aaatgggtga gggggagctc
      721 tccccatagc tgggctgcgg cccaacccca ccccctcagg ctatgccagg gggtgttgcc
      781 aggggcaccc gggcatcgcc agtctagccc actccttcat aaagccctcg catcccagga
      841 gcgagcagag ccagagcagg ttggagagga gacgcatcac ctccgctgct cgcgctagcg
      901 ctaccggtcg ccaccatgga agatgccaaa aacattaaga agggcccagc gccattctac
      961 ccactcgaag acgggaccgc cggcgagcag ctgcacaaag ccatgaagcg ctacgccctg
     1021 gtgcccggca ccatcgcctt taccgacgca catatcgagg tggacattac ctacgccgag
     1081 tacttcgaga tgagcgttcg gctggcagaa gctatgaagc gctatgggct gaatacaaac
     1141 catcggatcg tggtgtgcag cgagaatagc ttgcagttct tcatgcccgt gttgggtgcc
     1201 ctgttcatcg gtgtggctgt ggccccagct aacgacatct acaacgagcg cgagctgctg
     1261 aacagcatgg gcatcagcca gcccaccgtc gtattcgtga gcaagaaagg gctgcaaaag
     1321 atcctcaacg tgcaaaagaa gctaccgatc atacaaaaga tcatcatcat ggatagcaag
     1381 accgactacc agggcttcca aagcatgtac accttcgtga cttcccattt gccacccggc
     1441 ttcaacgagt acgacttcgt gcccgagagc ttcgaccggg acaaaaccat cgccctgatc
     1501 atgaacagta gtggcagtac cggattgccc aagggcgtag ccctaccgca ccgcaccgct
     1561 tgtgtccgat tcagtcatgc ccgcgacccc atcttcggca accagatcat ccccgacacc
     1621 gctatcctca gcgtggtgcc atttcaccac ggcttcggca tgttcaccac gctgggctac
     1681 ttgatctgcg gctttcgggt cgtgctcatg taccgcttcg aggaggagct attcttgcgc
     1741 agcttgcaag actataagat tcaatctgcc ctgctggtgc ccacactatt tagcttcttc
     1801 gctaagagca ctctcatcga caagtacgac ctaagcaact tgcacgagat cgccagcggc
     1861 ggggcgccgc tcagcaagga ggtaggtgag gccgtggcca aacgcttcca cctaccaggc
     1921 atccgccagg gctacggcct gacagaaaca accagcgcca ttctgatcac ccccgaaggg
     1981 gacgacaagc ctggcgcagt aggcaaggtg gtgcccttct tcgaggctaa ggtggtggac
     2041 ttggacaccg gtaagacact gggtgtgaac cagcgcggcg agctgtgcgt ccgtggcccc
     2101 atgatcatga gcggctacgt taacaacccc gaggctacaa acgctctcat cgacaaggac
     2161 ggctggctgc acagcggcga catcgcctac tgggacgagg acgagcactt cttcatcgtg
     2221 gaccggctga agagcctgat caaatacaag ggctaccagg tagccccagc cgaactggag
     2281 agcatcctgc tgcaacaccc caacatcttc gacgccgggg tcgccggcct gcccgacgac
     2341 gatgccggcg agctgcccgc cgcagtcgtc gtgctggaac acggtaaaac catgaccgag
     2401 aaggagatcg tggactatgt ggccagccag gttacaaccg ccaagaagct gcgcggtggt
     2461 gttgtgttcg tggacgaggt gcctaaagga ctgaccggca agttggacgc ccgcaagatc
     2521 cgcgagattc tcattaaggc caagaagggc ggcaagatcg ccgtgtaaga attctgcctc
     2581 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     2641 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     2701 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2761 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggggc ctcgagcgcg
     2821 tgtgaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     2881 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     2941 gagcgagcgc gcagctgcct gcaggatcgg atgccgggac cgacgagtgc agaggcgtgc
     3001 aagcgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3061 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3121 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3181 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3241 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3301 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3361 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3421 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3481 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3541 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3601 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3661 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3721 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3781 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3841 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3901 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3961 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4021 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4081 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4141 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4201 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4261 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4321 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4381 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4441 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4501 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4561 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4621 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4681 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4741 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4801 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4861 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4921 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4981 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5041 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5101 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5161 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5221 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5281 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5341 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5401 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     5461 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     5521 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5581 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5641 aattggagat cggtacttcg cgaatgcgtc gagat
//