LOCUS       62056_22745        6233 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 6233)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6233
                     /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        1378..1663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer;
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1665..1942
                     /label=chicken beta-actin promoter
     CDS             2209..3237
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="VSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             3238..3264
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    3298..3505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3514..3654
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3729..4184
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4466..4570
                     /label=AmpR promoter
     CDS             4571..5428
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5602..6190
                     /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 ctcgagctcc ctctgagccg 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 caccgcgtct accgttgcat cggtcgtttt
      781 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac
      841 cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
      901 ttttagcgcg tgcgccaatt ctgcagacaa atggggtacc ctccatcact aggggttcct
      961 gcggccgcac gcgtgagggc ctatttccca tgattccttc atatttgcat atacgataca
     1021 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     1081 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     1141 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     1201 tcttgtggaa aggacgaaac accggaagag cgagctcttc tgttttagag ctagaaatag
     1261 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     1321 ttttctagag gtaccaggtc ttgaaaggag tgggaattgg ctccggtgcc cgtcagtcgt
     1381 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     1441 gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     1501 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     1561 caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat gggactttcc
     1621 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac
     1681 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat
     1741 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg
     1801 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca
     1861 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     1921 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     1981 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     2041 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
     2101 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
     2161 tttttttcag gttggaccgg tgccaccatg cccaagaaga agaggaaggt gtccaatctc
     2221 ctgactgttc accagaacct ccctgcgctg ccagtagatg ccactagcga tgaggtcagg
     2281 aaaaatctca tggatatgtt tagggataga caggcgtttt ctgaacacac ctggaaaatg
     2341 ctgcttagcg tgtgccgatc ctgggcagcc tggtgtaagc tgaacaatcg caaatggttc
     2401 cccgccgagc cggaggacgt gcgcgattac ctgctgtatc tccaggcaag agggctggct
     2461 gtcaagacta tccagcagca cttgggccaa ctgaatatgc tgcatcgacg cagcgggctc
     2521 ccccggccta gcgattcaaa cgcagtctcc cttgttatga ggagaattag aaaggaaaac
     2581 gtagatgcgg gtgagagggc taagcaggct ctcgcttttg agcggactga tttcgaccag
     2641 gtcagatccc tgatggagaa cagcgatcgg tgccaggaca tcaggaacct cgcatttctg
     2701 ggaattgcat ataacacact tctgcgcata gctgagatcg cccggatcag agtgaaagac
     2761 atcagtcgaa cggacggcgg ccggatgctt attcatattg gacgcacaaa gacattggtc
     2821 agcaccgctg gcgttgaaaa ggccttgtcc ctgggcgtaa cgaagctggt ggaaagatgg
     2881 atctcagtgt ccggcgtggc tgacgaccct aataattact tgttctgtcg agtgagaaaa
     2941 aacggagtcg ccgcgccctc tgccaccagc caattgagta cacgggccct tgaagggatc
     3001 tttgaggcaa cccaccgact catatacgga gccaaggatg acagtggcca gaggtatctc
     3061 gcctggtcag gtcattctgc tagggtgggg gccgcacgag acatggcgcg ggcaggagtc
     3121 tccataccag agattatgca agctggaggt tggacaaatg tgaacatcgt tatgaactat
     3181 atccgcaatc ttgactctga aaccggggcc atggtgagac tgctcgaaga tggtgactac
     3241 ccatacgatg ttccagatta cgcttaagaa ttcctagagc tcgctgatca gcctcgactg
     3301 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3361 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3421 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3481 aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca
     3541 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3601 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     3661 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     3721 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3781 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     3841 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3901 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     3961 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4021 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     4081 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     4141 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     4201 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     4261 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4321 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     4381 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     4441 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4501 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4561 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4621 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4681 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4741 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4801 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4861 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4921 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4981 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5041 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5101 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5161 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5221 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5281 gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5341 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5401 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5461 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5521 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gct
//
