LOCUS       sequence-467619-        3347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contain AAV ITRs, lacZ expressing for blue-white selection for 
            golden-gate reaction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3347)
  AUTHORS   Luo Y, Lin L, Bolund L, Sorensen CB
  TITLE     Efficient construction of rAAV-based gene targeting vectors by 
            Golden Gate cloning.
  JOURNAL   Biotechniques. 2014 May 1;56(5):263-8. doi: 10.2144/000114169. 
            eCollection 2014 May.
  PUBMED    24806227
REFERENCE   2  (bases 1 to 3347)
  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"; doi: "10.2144/000114169";
            journalName: "Biotechniques"; date: "2014-05-1- 1"; volume: "56"; 
            issue: "5"; pages: "263-8"
COMMENT     Sequence Label: pGolden-AAV
FEATURES             Location/Qualifiers
     source          1..3347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    184..205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(220..225,226..243,244..250)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    258..274
                     /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     263..285
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     282..298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     282..298
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             294..848
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGAGLDRR
                     PQEPLVMELATPSLRARSLTEAGRPKVARRPGFARAASVSERARSCLQGRLMRYFLLTH
                     LCGISHRIRQSNHSTRPVAAH"
     primer_bind     317..337
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        319..337
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(384..403)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(385..403)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(413..430)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(413..429)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(422..444)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     repeat_region   610..750
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   610..739
                     /label=AAV2 ITR
     rep_origin      825..1280
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(912..931)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1122..1143
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1297..1316)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1416..1438
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1476..1494)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1562..1666
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1667..1735,1736..2527)
                     /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(1885..1904)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2698..3286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3187..3206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgccg ccagagacca ccggtggaaa gcgggcagtg agcgcaacgc
      181 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      241 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      301 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg aggacctcga
      361 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
      421 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
      481 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
      541 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgcc ggtctcgata
      601 ggcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc
      661 tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag
      721 tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc
      781 atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg
      841 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      901 cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      961 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1021 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1081 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1141 tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     1201 ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1261 aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata
     1321 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1381 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1441 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1501 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1561 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     1621 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     1681 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     1741 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     1801 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     1861 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     1921 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     1981 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2041 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     2101 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2161 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2221 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2281 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2341 tggctggttt attgctgata aatctggagc cggtgagcgt ggctcgcgcg gtatcattgc
     2401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2521 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     2581 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     2641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2821 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//
