LOCUS       62056_22735        6238 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 6238)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6238
                     /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        340..415
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1383..1668
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1670..1947
                     /label=chicken beta-actin promoter
     CDS             2214..3242
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="VSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             3243..3269
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    3303..3510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3519..3659
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3734..4189
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4471..4575
                     /label=AmpR promoter
     CDS             4576..5433
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5607..6195
                     /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 gaaacaccgc gtctaccgtt gcatcggtcg ttttagagct agaaatagca
      361 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      421 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc
      481 aattctgcag acaaatggct ctagagaggg cctatttccc atgattcctt catatttgca
      541 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      601 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      661 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      721 ggctttatat atcttgtgga aaggacgaaa caccggagac ctattcgcct taaggtctcg
      781 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      841 ggcaccgagt cggtgctttt ttgttttaga gctagaaata gcaagttaaa ataaggctag
      901 tccgttttta gcgcgtgcgc caattctgca gacaaatggg gtaccctcca tcactagggg
      961 ttcctgcggc cgcacgcgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     1021 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     1081 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     1141 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     1201 atatatcttg tggaaaggac gaaacaccgg aagagcgagc tcttctgttt tagagctaga
     1261 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     1321 gctttttttc tagaggtacc aggtcttgaa aggagtggga attggctccg gtgcccgtca
     1381 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1441 ttgacgtcaa tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1501 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1561 gacgtcaatg acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac
     1621 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
     1681 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     1741 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca
     1801 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     1861 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     1921 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     1981 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     2041 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg
     2101 tttaagggat ggttggttgg tggggtatta atgtttaatt acctggagca cctgcctgaa
     2161 atcacttttt ttcaggttgg accggtgcca ccatgcccaa gaagaagagg aaggtgtcca
     2221 atctcctgac tgttcaccag aacctccctg cgctgccagt agatgccact agcgatgagg
     2281 tcaggaaaaa tctcatggat atgtttaggg atagacaggc gttttctgaa cacacctgga
     2341 aaatgctgct tagcgtgtgc cgatcctggg cagcctggtg taagctgaac aatcgcaaat
     2401 ggttccccgc cgagccggag gacgtgcgcg attacctgct gtatctccag gcaagagggc
     2461 tggctgtcaa gactatccag cagcacttgg gccaactgaa tatgctgcat cgacgcagcg
     2521 ggctcccccg gcctagcgat tcaaacgcag tctcccttgt tatgaggaga attagaaagg
     2581 aaaacgtaga tgcgggtgag agggctaagc aggctctcgc ttttgagcgg actgatttcg
     2641 accaggtcag atccctgatg gagaacagcg atcggtgcca ggacatcagg aacctcgcat
     2701 ttctgggaat tgcatataac acacttctgc gcatagctga gatcgcccgg atcagagtga
     2761 aagacatcag tcgaacggac ggcggccgga tgcttattca tattggacgc acaaagacat
     2821 tggtcagcac cgctggcgtt gaaaaggcct tgtccctggg cgtaacgaag ctggtggaaa
     2881 gatggatctc agtgtccggc gtggctgacg accctaataa ttacttgttc tgtcgagtga
     2941 gaaaaaacgg agtcgccgcg ccctctgcca ccagccaatt gagtacacgg gcccttgaag
     3001 ggatctttga ggcaacccac cgactcatat acggagccaa ggatgacagt ggccagaggt
     3061 atctcgcctg gtcaggtcat tctgctaggg tgggggccgc acgagacatg gcgcgggcag
     3121 gagtctccat accagagatt atgcaagctg gaggttggac aaatgtgaac atcgttatga
     3181 actatatccg caatcttgac tctgaaaccg gggccatggt gagactgctc gaagatggtg
     3241 actacccata cgatgttcca gattacgctt aagaattcct agagctcgct gatcagcctc
     3301 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3361 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3421 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3481 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     3541 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3601 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3661 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3721 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3781 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     3841 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3901 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     3961 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4021 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4081 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4141 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4201 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4261 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4321 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4381 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4441 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4501 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4561 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4621 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4681 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4741 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4801 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4861 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4921 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4981 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5041 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5101 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5161 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5221 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5281 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5341 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5401 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5461 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5521 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5581 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5641 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5701 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5761 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5821 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5881 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5941 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6001 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6061 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6121 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6181 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgct
//
