LOCUS       62056_22730        6235 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        70..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        337..412
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1380..1665
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1667..1944
                     /label=chicken beta-actin promoter
     CDS             2211..3239
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="VSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             3240..3266
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    3300..3507
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3516..3656
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3731..4186
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4468..4572
                     /label=AmpR promoter
     CDS             4573..5430
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5604..6192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggccttttgc tcacatgcgc cattctcgag catgcgccat tctcgagcat gctcgagaat
       61 ggcgcatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
      121 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
      181 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
      241 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
      301 tggaaaggac gaaacaccgg gtcttcgaga agacctgttt tagagctaga aatagcaagt
      361 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gcttttttgt
      421 tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc gtgcgccaat
      481 tctgcagaca aatggctcta gagagggcct atttcccatg attccttcat atttgcatat
      541 acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt
      601 agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt
      661 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc
      721 tttatatatc ttgtggaaag gacgaaacac cggagaccta ttcgccttaa ggtctcggtt
      781 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
      841 accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc
      901 gtttttagcg cgtgcgccaa ttctgcagac aaatggggta ccctccatca ctaggggttc
      961 ctgcggccgc acgcgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     1021 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     1081 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     1141 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     1201 tatcttgtgg aaaggacgaa acaccggaag agcgagctct tctgttttag agctagaaat
     1261 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     1321 ttttttctag aggtaccagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtc
     1381 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     1441 acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1501 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1561 gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt atgggacttt
     1621 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
     1681 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     1741 attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccaggc
     1801 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
     1861 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
     1921 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
     1981 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
     2041 agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt
     2101 aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc
     2161 actttttttc aggttggacc ggtgccacca tgcccaagaa gaagaggaag gtgtccaatc
     2221 tcctgactgt tcaccagaac ctccctgcgc tgccagtaga tgccactagc gatgaggtca
     2281 ggaaaaatct catggatatg tttagggata gacaggcgtt ttctgaacac acctggaaaa
     2341 tgctgcttag cgtgtgccga tcctgggcag cctggtgtaa gctgaacaat cgcaaatggt
     2401 tccccgccga gccggaggac gtgcgcgatt acctgctgta tctccaggca agagggctgg
     2461 ctgtcaagac tatccagcag cacttgggcc aactgaatat gctgcatcga cgcagcgggc
     2521 tcccccggcc tagcgattca aacgcagtct cccttgttat gaggagaatt agaaaggaaa
     2581 acgtagatgc gggtgagagg gctaagcagg ctctcgcttt tgagcggact gatttcgacc
     2641 aggtcagatc cctgatggag aacagcgatc ggtgccagga catcaggaac ctcgcatttc
     2701 tgggaattgc atataacaca cttctgcgca tagctgagat cgcccggatc agagtgaaag
     2761 acatcagtcg aacggacggc ggccggatgc ttattcatat tggacgcaca aagacattgg
     2821 tcagcaccgc tggcgttgaa aaggccttgt ccctgggcgt aacgaagctg gtggaaagat
     2881 ggatctcagt gtccggcgtg gctgacgacc ctaataatta cttgttctgt cgagtgagaa
     2941 aaaacggagt cgccgcgccc tctgccacca gccaattgag tacacgggcc cttgaaggga
     3001 tctttgaggc aacccaccga ctcatatacg gagccaagga tgacagtggc cagaggtatc
     3061 tcgcctggtc aggtcattct gctagggtgg gggccgcacg agacatggcg cgggcaggag
     3121 tctccatacc agagattatg caagctggag gttggacaaa tgtgaacatc gttatgaact
     3181 atatccgcaa tcttgactct gaaaccgggg ccatggtgag actgctcgaa gatggtgact
     3241 acccatacga tgttccagat tacgcttaag aattcctaga gctcgctgat cagcctcgac
     3301 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3361 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3421 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3481 ggaagagaat agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc
     3541 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3601 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3661 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3721 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3781 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3841 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3901 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     3961 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4021 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4081 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4141 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4201 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4261 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4321 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4381 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4441 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4501 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4561 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4621 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4681 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4741 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4801 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4861 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4921 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4981 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5041 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5101 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5161 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5221 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5281 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5341 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5401 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5461 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5521 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5581 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5641 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5701 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5761 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5821 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5881 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5941 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6001 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6061 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6121 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6181 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgct
//
