LOCUS       40924_47048        5303 bp DNA     circular SYN 11-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5303)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5303)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        156..396
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        423..498
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        532..979
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1017..1733
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    2050..2526
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   2566..2706
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      2781..3236
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3518..3622
                     /label=AmpR promoter
     CDS             3623..4480
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4654..5242
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgaggg cctatttccc atgattcctt
      181 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
      241 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
      301 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
      361 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccggaaga gcgagctctt
      421 ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa
      481 gtggcaccga gtcggtgctt tttttctaga ctgcagaggg ccctgcgtat gagtgcaagt
      541 gggttttagg accaggatga ggcggggtgg gggtgcctac ctgacgaccg accccgaccc
      601 actggacaag cacccaaccc ccattcccca aattgcgcat cccctatcag agagggggag
      661 gggaaacagg atgcggcgag gcgcgtgcgc actgccagct tcagcaccgc ggacagtgcc
      721 ttcgcccccg cctggcggcg cgcgccaccg ccgcctcagc actgaaggcg cgctgacgtc
      781 actcgccggt cccccgcaaa ctccccttcc cggccacctt ggtcgcgtcc gcgccgccgc
      841 cggcccagcc ggaccgcacc acgcgaggcg cgagataggg gggcacgggc gcgaccatct
      901 gcgctgcggc gccggcgact cagcgctgcc tcagtctgcg gtgggcagcg gaggagtcgt
      961 gtcgtgcctg agagcgcagt cgagaaggta ccggatcctc tagagtcgac gccaccatgg
     1021 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1081 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1141 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1201 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1261 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1321 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1381 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1441 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1501 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1561 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1621 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1681 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtccggac
     1741 tcagatctcg agaggaggag gaggagacag acagcaggat gccccacctc gacagccccg
     1801 gcagctccca gccgagacgc tccttcctct caagggtgat cagggcagcg ctaccgttgc
     1861 agctgcttct gctgctgctg ctgctcctgg cctgcctgct acctgcctct gaagatgact
     1921 acagctgcac ccaggccaac aactttgccc gatccttcta ccccatgctg cggtacacca
     1981 acgggccacc tcccacctag gaattcgata tcaagcttat cgataccgag cgctgctcga
     2041 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     2101 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     2161 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     2221 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     2281 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     2341 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     2401 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     2461 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     2521 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     2581 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2641 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     2701 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2761 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3001 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3061 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3121 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3181 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     3241 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3301 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3361 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3421 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3481 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3541 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3601 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     3661 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3721 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3781 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3841 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3901 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3961 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4021 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4081 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4141 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4201 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4261 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4321 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4381 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4441 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4501 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4561 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4621 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4681 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4741 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4801 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4861 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4921 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4981 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5041 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5101 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5161 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5221 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5281 ttgctggcct tttgctcaca tgt
//
