LOCUS       Exported                6036 bp ds-DNA     circular SYN 30-AUG-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEF1a-GFP-gRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Aug 30, 2017 from SnapGene 3.2.1
            http://www.snapgene.com
COMMENT     Note: stop codon is in IVS sequence.
FEATURES             Location/Qualifiers
     source          1..6036
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        201..1458
                     /note="EF1a Promoter"
     CDS             1474..2187
                     /codon_start=1
                     /note="GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    2188..2250
                     /note="2a"
     CDS             2251..2844
                     /codon_start=1
                     /note="Puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     intron          2848..3145
                     /note="IVS"
     polyA_signal    3187..3425
                     /note="SV40 polyA"
     promoter        3481..3779
                     /note="U6 Promoter"
     misc_feature    3780..3836
                     /note="gRNA Scaffold"
     terminator      3837..3842
                     /note="Poly T terminator"
     rep_origin      4266..4885
                     /note="pBR322 Origin"
     CDS             complement(5040..5900)
                     /codon_start=1
                     /note="Ampicillin"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        5942..5970
                     /note="Amp promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg tgcaaagatg gataaagttt taaacagaga ggaatctttg
      241 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      301 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      361 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      421 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      481 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
      541 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccactg gctgcagtac
      601 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
      661 caaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
      721 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
      781 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
      841 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
      901 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
      961 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     1021 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     1081 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     1141 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     1201 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     1261 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     1321 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     1381 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     1441 ccatttcagg tgtcgtgacg tacggccacc atggtgagca agggcgagga gctgttcacc
     1501 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1561 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1621 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1681 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1741 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1801 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1861 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1921 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1981 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2041 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2101 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2161 actctcggca tggacgagct gtacaagggc agtggagagg gcagaggaag tctgctaaca
     2221 tgcggtgacg tcgaggagaa tcctggccca accgagtaca agcccacggt gcgcctcgcc
     2281 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     2341 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     2401 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     2461 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     2521 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     2581 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtgtcg
     2641 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     2701 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     2761 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     2821 tgcatgaccc gcaagcccgg tgcctgatgg aattaattcg ctgtctgcga gggccagctg
     2881 ttggggtgag tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc
     2941 caaaaacgag gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc
     3001 gtccatctgg tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga
     3061 gatctggcca tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc
     3121 cactcccagg tccaactgca ggtcgattta aatgggaatt ccacagcggc cgcatgtcta
     3181 gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3241 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3301 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3361 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaatgcg
     3421 tcgagtgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3481 tgtacaaaaa agcaggcttt aaaggaacca attcagtcga ctggatccgg taccaaggtc
     3541 gggcaggaag agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     3601 gttagagaga taattagaat taatttgact gtaaacacaa agatattagt acaaaatacg
     3661 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     3721 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatctta
     3781 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     3841 ttaccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     3901 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3961 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4021 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4081 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4441 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     4501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4681 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6001 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
