EJ2GFP-puro

LOCUS       EJ2GFP-puro               7636 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    EJ2GFP-puro
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7636)
  AUTHORS   Bennardo N, Cheng A, Huang N, Stark JM
  TITLE     Alternative-NHEJ is a mechanistically distinct pathway of mammalian 
            chromosome break repair.
  JOURNAL   PLoS Genet. 2008 Jun 27;4(6):e1000110. doi: 
            10.1371/journal.pgen.1000110.
  PUBMED    18584027
REFERENCE   2  (bases 1 to 7636)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7636
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        29..526
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(64..87)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     459..478
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             545..1144
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(545..564)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1041..1061
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        1788..2167
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2170..2445
                     /label=chicken beta-actin promoter
     intron          2446..3457
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3381..3403
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3465..3484
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3546..3566
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3588..3608
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      3857..3866
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3863..4582
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3908..3929)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4169..4188)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4516..4537
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4787..4806)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4852..4907
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(4906..4925)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(5263..5281)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5264..5281)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5362..5381)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5481..5503
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5541..5559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5627..5731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5732..6592
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5950..5969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6763..7351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7252..7271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7505..7522
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7609..7628
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7609..7627
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agatctgata tcatcgatga attctaccgg gtaggggagg cgcttttccc aaggcagtct
       61 ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct
      121 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
      181 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
      241 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
      301 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
      361 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
      421 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
      481 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgaccg atccagccgc
      541 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc
      601 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc
      661 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
      721 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
      781 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
      841 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
      901 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
      961 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     1021 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     1081 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     1141 ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc
     1201 cgaccgaagc cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgcg
     1261 ggggacacac cgaacacgcc gaccctgctg aacacgcggc gcagttcggt gcccaggagc
     1321 ggatcgaaat tgatgatcta ttaaacaata aagatgtcca ctaaaatgga agtttttcct
     1381 gtcatacttt gttaagaagg gtgagaacag agtacctaca ttttgaatgg aaggattgga
     1441 gctacggggg tgggggtggg gtgggattag ataaatgcct gctctttact gaaggctctt
     1501 tactattgct ttatgataat gtttcatagt tggatatcat aatttaaaca agcaaaacca
     1561 aattaagggc cagctcattc ctcccactca tgatctatag atctatagat ctctcgtggg
     1621 atcattgttt ttctcttgat tcccactttg tggttctaag tactgtggtt tccaaatgtg
     1681 tcagtttcat agcctgaaga acgagatcag cagcctctgt tccacataca cttcattctc
     1741 agtattgttt tgccaagttc taattccatc agaagcttca gctgctcgac attgattatt
     1801 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     1861 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     1921 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     1981 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     2041 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     2101 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     2161 taccatgggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     2221 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     2281 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     2341 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     2401 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgttg
     2461 ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc cgccccggct ctgactgacc
     2521 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
     2581 ttggtttaat gacggctcgt ttcttttctg tggctgcgtg aaagccttaa agggctccgg
     2641 gagggccctt tgtgcggggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
     2701 gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg
     2761 ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     2821 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     2881 gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca cccccctccc cgagttgctg
     2941 agcacggccc ggcttcgggt gcggggctcc gtgcggggcg tggcgcgggg ctcgccgtgc
     3001 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     3061 agggctcggg ggaggggcgc ggcggccccg gagcgccggc ggctgtcgag gcgcggcgag
     3121 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     3181 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     3241 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     3301 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     3361 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     3421 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     3481 tgtgctgtct catcattttg gcaaagaatt aattcagatc cgccgccact atgggatcaa
     3541 gatcgccaaa aaagaagaga aaggtgccga agaagcatgc agcaccacca aaaaaaaaac
     3601 gaaaagtaga agacccacga ggcaacacca gcggcgtgct gagcaccccc aaggccaaga
     3661 gggccaagca cccccccggc accgagaagc ccaggagcag gagccagagc gagcagcccg
     3721 ccacctgccc catctgctac gccgtgatca ggcagagcag gaacctgagg aggcacctgg
     3781 agctgaggca cttcgccaag cccgcctagg gataacaggg taattagatg acaagcccgg
     3841 cgtggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     3901 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3961 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4021 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4081 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4141 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     4201 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     4261 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     4321 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     4381 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     4441 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     4501 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     4561 catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca
     4621 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     4681 aaaatgaatg caattgttgt tgttaattca ctcctcaggt gcaggctgcc tatcagaagg
     4741 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4801 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4861 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4921 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4981 atgccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     5041 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     5101 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     5161 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     5221 ttatgaagat ccctcgacct gcagcccaag ctggcgtcga gttctatagt gtcacctaaa
     5281 tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta acgacaatat
     5341 gtccatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5401 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5461 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5521 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6061 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6121 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6181 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6241 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6421 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6541 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6901 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7141 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7381 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7441 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7501 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7561 cgcgcgttgg ccgattcatt aatgcaggtt aacctggctt atcgaaatta atacgactca
     7621 ctatagggag accggc
//