LOCUS       Exported                7283 bp DNA     circular SYN 26-MAR-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    rAAV2/MG1.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7283)
  AUTHORS   Luo Lab
  TITLE     Direct Submission
  JOURNAL   Exported Mar 26, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..39
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     31..48
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     32..48
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     58..77
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        58..76
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     105..121
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     106..122
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             177..2042
                     /codon_start=1
                     /label=Rep2
                     /translation="MPGFYEIVIKVPSDLDEHLPGISDSFVNWVAEKEWELPPDSDMDL
                     NLIEQAPLTVAEKLQRDFLTEWRRVSKAPEALFFVQFEKGESYFHMHVLVETTGVKSMV
                     LGRFLSQIREKLIQRIYRGIEPTLPNWFAVTKTRNGAGGGNKVVDECYIPNYLLPKTQP
                     ELQWAWTNMEQYLSACLNLTERKRLVAQHLTHVSQTQEQNKENQNPNSDAPVIRSKTSA
                     RYMELVGWLVDKGITSEKQWIQEDQASYISFNAASNSRSQIKAALDNAGKIMSLTKTAP
                     DYLVGQQPVEDISSNRIYKILELNGYDPQYAASVFLGWATKKFGKRNTIWLFGPATTGK
                     TNIAEAIAHTVPFYGCVNWTNENFPFNDCVDKMVIWWEEGKMTAKVVESAKAILGGSKV
                     RVDQKCKSSAQIDPTPVIVTSNTNMCAVIDGNSTTFEHQQPLQDRMFKFELTRRLDHDF
                     GKVTKQEVKDFFRWAKDHVVEVEHEFYVKKGGAKKRPAPSDADISEPKRVRESVAQPST
                     SDAEASINYADRYQNKCSRHVGMNLMLFPCRQCERMNQNSNICFTHGQKDCLECFPVSE
                     SQPVSVVKKAYQKLCYIHHIMGKVPDACTACDLVNVDLDDCIFEQ"
     CDS             2059..4290
                     /codon_start=1
                     /label=AAV-MG1.2 Capsid
                     /label=AAV9 Capsid
                     /translation="MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGL
                     VLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERL
                     KEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSG
                     AQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVG
                     SSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPW
                     GYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTS
                     TVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEY
                     FPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSRTINGSGQNQ
                     QTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSL
                     MNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATES
                     YGQVATNHQSATEPPKTTSAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGN
                     FHPSPLMGGFGMKHPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWEL
                     QKENSKRWNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL"
     misc_difference 3403..3405
                     /label=K449R
     misc_feature    3820..3822
                     /label=Q588T
     misc_feature    3823..3843
                     /label=Insertion
     primer_bind     complement(4527..4547)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4527..4545)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4566..4582)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4566..4582)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4579..4601)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4590..4606
                     /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(4614..4644)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4659..4680
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(4797..4814)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4968..5556)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5048..5067)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5727..6587)
                     /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     6350..6369
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6588..6692)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6718..7173)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6855..6876)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7067..7086
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa
       61 tacgactcac tatagggcga attgggtacc gggccccccc tcgatcgagg tcgacggtat
      121 cgggggagct cgcagggtct ccattttgaa gcgggaggtt tgaacgcgca gccgccatgc
      181 cggggtttta cgagattgtg attaaggtcc ccagcgacct tgacgagcat ctgcccggca
      241 tttctgacag ctttgtgaac tgggtggccg agaaggaatg ggagttgccg ccagattctg
      301 acatggatct gaatctgatt gagcaggcac ccctgaccgt ggccgagaag ctgcagcgcg
      361 actttctgac ggaatggcgc cgtgtgagta aggccccgga ggctcttttc tttgtgcaat
      421 ttgagaaggg agagagctac ttccacatgc acgtgctcgt ggaaaccacc ggggtgaaat
      481 ccatggtttt gggacgtttc ctgagtcaga ttcgcgaaaa actgattcag agaatttacc
      541 gcgggatcga gccgactttg ccaaactggt tcgcggtcac aaagaccaga aatggcgccg
      601 gaggcgggaa caaggtggtg gatgagtgct acatccccaa ttacttgctc cccaaaaccc
      661 agcctgagct ccagtgggcg tggactaata tggaacagta tttaagcgcc tgtttgaatc
      721 tcacggagcg taaacggttg gtggcgcagc atctgacgca cgtgtcgcag acgcaggagc
      781 agaacaaaga gaatcagaat cccaattctg atgcgccggt gatcagatca aaaacttcag
      841 ccaggtacat ggagctggtc gggtggctcg tggacaaggg gattacctcg gagaagcagt
      901 ggatccagga ggaccaggcc tcatacatct ccttcaatgc ggcctccaac tcgcggtccc
      961 aaatcaaggc tgccttggac aatgcgggaa agattatgag cctgactaaa accgcccccg
     1021 actacctggt gggccagcag cccgtggagg acatttccag caatcggatt tataaaattt
     1081 tggaactaaa cgggtacgat ccccaatatg cggcttccgt ctttctggga tgggccacga
     1141 aaaagttcgg caagaggaac accatctggc tgtttgggcc tgcaactacc gggaagacca
     1201 acatcgcgga ggccatagcc cacactgtgc ccttctacgg gtgcgtaaac tggaccaatg
     1261 agaactttcc cttcaacgac tgtgtcgaca agatggtgat ctggtgggag gaggggaaga
     1321 tgaccgccaa ggtcgtggag tcggccaaag ccattctcgg aggaagcaag gtgcgcgtgg
     1381 accagaaatg caagtcctcg gcccagatag acccgactcc cgtgatcgtc acctccaaca
     1441 ccaacatgtg cgccgtgatt gacgggaact caacgacctt cgaacaccag cagccgttgc
     1501 aagaccggat gttcaaattt gaactcaccc gccgtctgga tcatgacttt gggaaggtca
     1561 ccaagcagga agtcaaagac tttttccggt gggcaaagga tcacgtggtt gaggtggagc
     1621 atgaattcta cgtcaaaaag ggtggagcca agaaaagacc cgcccccagt gacgcagata
     1681 taagtgagcc caaacgggtg cgcgagtcag ttgcgcagcc atcgacgtca gacgcggaag
     1741 cttcgatcaa ctacgcagac aggtaccaaa acaaatgttc tcgtcacgtg ggcatgaatc
     1801 tgatgctgtt tccctgcaga caatgcgaga gaatgaatca gaattcaaat atctgcttca
     1861 ctcacggaca gaaagactgt ttagagtgct ttcccgtgtc agaatctcaa cccgtttctg
     1921 tcgtcaaaaa ggcgtatcag aaactgtgct acattcatca tatcatggga aaggtgccag
     1981 acgcttgcac tgcctgcgat ctggtcaatg tggatttgga tgactgcatc tttgaacaat
     2041 aaatgattta aatcaggtat ggctgccgat ggttatcttc cagattggct cgaggacaac
     2101 cttagtgaag gaattcgcga gtggtgggct ttgaaacctg gagcccctca acccaaggca
     2161 aatcaacaac atcaagacaa cgctcgaggt cttgtgcttc cgggttacaa ataccttgga
     2221 cccggcaacg gactcgacaa gggggagccg gtcaacgcag cagacgcggc ggccctcgag
     2281 cacgacaagg cctacgacca gcagctcaag gccggagaca acccgtacct caagtacaac
     2341 cacgccgacg ccgagttcca ggagcggctc aaagaagata cgtcttttgg gggcaacctc
     2401 gggcgagcag tcttccaggc caaaaagagg cttcttgaac ctcttggtct ggttgaggaa
     2461 gcggctaaga cggctcctgg aaagaagagg cctgtagagc agtctcctca ggaaccggac
     2521 tcctccgcgg gtattggcaa atcgggtgca cagcccgcta aaaagagact caatttcggt
     2581 cagactggcg acacagagtc agtcccagac cctcaaccaa tcggagaacc tcccgcagcc
     2641 ccctcaggtg tgggatctct tacaatggct tcaggtggtg gcgcaccagt ggcagacaat
     2701 aacgaaggtg ccgatggagt gggtagttcc tcgggaaatt ggcattgcga ttcccaatgg
     2761 ctgggggaca gagtcatcac caccagcacc cgaacctggg ccctgcccac ctacaacaat
     2821 cacctctaca agcaaatctc caacagcaca tctggaggat cttcaaatga caacgcctac
     2881 ttcggctaca gcaccccctg ggggtatttt gacttcaaca gattccactg ccacttctca
     2941 ccacgtgact ggcagcgact catcaacaac aactggggat tccggcctaa gcgactcaac
     3001 ttcaagctct tcaacattca ggtcaaagag gttacggaca acaatggagt caagaccatc
     3061 gccaataacc ttaccagcac ggtccaggtc ttcacggact cagactatca gctcccgtac
     3121 gtgctcgggt cggctcacga gggctgcctc ccgccgttcc cagcggacgt tttcatgatt
     3181 cctcagtacg ggtatctgac gcttaatgat ggaagccagg ccgtgggtcg ttcgtccttt
     3241 tactgcctgg aatatttccc gtcgcaaatg ctaagaacgg gtaacaactt ccagttcagc
     3301 tacgagtttg agaacgtacc tttccatagc agctacgctc acagccaaag cctggaccga
     3361 ctaatgaatc cactcatcga ccaatacttg tactatctct ctagaactat taacggttct
     3421 ggacagaatc aacaaacgct aaaattcagt gtggccggac ccagcaacat ggctgtccag
     3481 ggaagaaact acatacctgg acccagctac cgacaacaac gtgtctcaac cactgtgact
     3541 caaaacaaca acagcgaatt tgcttggcct ggagcttctt cttgggctct caatggacgt
     3601 aatagcttga tgaatcctgg acctgctatg gccagccaca aagaaggaga ggaccgtttc
     3661 tttcctttgt ctggatcttt aatttttggc aaacaaggta ccggtagaga caacgtggat
     3721 gcggacaaag tcatgataac caacgaagaa gaaattaaaa ctactaaccc ggtagcaacg
     3781 gagtcctatg gacaagtggc cacaaaccac cagagtgcca ctgagcctcc gaagactacg
     3841 tctgcacagg cgcagaccgg ttgggttcaa aaccaaggaa tacttccggg tatggtttgg
     3901 caggacagag atgtgtacct gcaaggaccc atttgggcca aaattcctca cacggacggc
     3961 aactttcacc cttctccgct gatgggaggg tttggaatga agcacccgcc tcctcagatc
     4021 ctcatcaaaa acacacctgt acctgcggat cctccaacgg ccttcaacaa ggacaagctg
     4081 aactctttca tcacccagta ttctactggc caagtcagcg tggagatcga gtgggagctg
     4141 cagaaggaaa acagcaagcg ctggaacccg gagatccagt acacttccaa ctattacaag
     4201 tctaataatg ttgaatttgc tgttaatact gaaggtgtat atagtgaacc ccgccccatt
     4261 ggcaccagat acctgactcg taatctgtaa gtttaaacct gcaggactag aggtcctgta
     4321 ttagaggtca cgtgagtgtt ttgcgacatt ttgcgacacc atgtggtcac gctgggtatt
     4381 taagcccgag tgagcacgca gggtctccat tttgaagcgg gaggtttgaa cgcgcagccg
     4441 ccaagccgaa ttctgcagat atccatcaca ctggcggccg ctcgactaga gcggccgcca
     4501 ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
     4561 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4621 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4681 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4741 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     4801 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4861 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4921 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4981 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5041 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5101 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5161 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5221 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5281 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5341 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5401 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5461 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5521 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5581 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5641 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5701 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5761 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5821 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     5881 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5941 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6001 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6061 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6121 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6181 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6241 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6301 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6361 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6421 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6481 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6541 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6601 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6661 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     6721 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     6781 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     6841 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6901 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6961 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     7021 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     7081 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     7141 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
     7201 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
     7261 gtgctgcaag gcgattaagt tgg
//
