LOCUS       Exported                5873 bp ds-DNA     circular SYN 15-JAN-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5-FRT-TO-eGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5873)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 19 Jan, 2016 from SnapGene Viewer 3.0.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     protein_bind    820..838
                     /dnas_title="tetO"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /dnas_title="tetO"
                     /gene="tetO"
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             990..1706
                     /dnas_title="enhanced GFP"
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT
                     LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN
                     GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYK"
     polyA_signal    2919..3143
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3427..3474
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3482..4504
                     /dnas_title="HygR"
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGDFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYH
                     WQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSE
                     AMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQS
                     LVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPST
                     RPRAKE"
     polyA_signal    4634..4755
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4804..4820)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    4828..4844
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4852..4882)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4918
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the
                     presence of cAMP."
     rep_origin      complement(5206..5794)
                     /direction=LEFT
                     /dnas_title="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5965..6825)
                     /dnas_title="AmpR"
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     promoter        complement(6826..6930)
                     /dnas_title="bla"
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             1716..2852
                     /note="NFATC2IP delMet del aa101-140"
                     /translation="AEPVGKRGRWSGGSGAGRGGRGGWGGRGRRPRAQRSPSRGTLDV
                     VSVDLVTDSDEEILEVATARGAADEVEVEPPEPPGPVASRDNSNSDSEGEDRRPALSL
                     LKLYPPGDEEEAELADSSGLYHEGSPSPGSPWKTKLRTKDKEEKKKTEFLDLDNSPLS
                     PPSPRTKSRTHTRALKKLSEVNKRLQDLRSCLSPKPPQGQEQQGQEDEVVLVEGPTLP
                     ETPRLFPLKIRCRADLVRLPLRMSEPLQSVVDHMATHLGVSPSRILLLFGETELSPTA
                     TPRTLKLGVADIIDCVVLTSSPEATETSQQLQLRVQGKEKHQTLEVSLSRDSPLKTLM
                     SHYEEAMGLSGRKLSFFFDGTKLSGRELPADLGMESGDLIEVWG"
     source          1..6961
                     /dnas_title="pcDNA5-FRT-TO-eGFP-NFATC2IP delM del101-140"
ORIGIN      
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca tggtgagcaa gggcgaggag ctgttcaccg
     1021 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     1081 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     1141 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     1201 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     1261 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1321 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1381 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1441 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1501 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1561 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1621 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1681 ctctcggcat ggacgagctg tacaagggCG CGCCAgcgga gcctgtgggg aagcggggcc
     1741 gctggtccgg aggtagcggt gccggccgag ggggtcgggg cggctggggc ggtcggggcc
     1801 ggcgtcctcg ggcccagcgg tctccatccc ggggcacgct ggacgtagtg tctgtggact
     1861 tggtcaccga cagcgatgag gaaattctgg aggtcgccac cgctcgcggt gccgcggacg
     1921 aggttgaggt ggagcccccg gagcccccgg ggccggtcgc gtcccgggat aacagcaaca
     1981 gtgacagcga aggggaggac aggcggcccg cactatccct cctgaaactc taccctccag
     2041 gggatgagga agaggcagag ctggcagatt cgagtggtct ctaccatgag ggctccccat
     2101 caccaggctc tccctggaag acaaagctga ggactaagga taaagaagag aagaaaaaga
     2161 cagagtttct ggatctggac aactctcctc tgtccccacc ttcaccaagg accaaaagca
     2221 gaacgcatac tcgggcactc aagaagttaa gtgaggtgaa caagcgcctc caggatctcc
     2281 gttcctgtct gagccccaag ccacctcagg gtcaagagca acagggccaa gaggatgaag
     2341 tggtcttggt ggaagggccc accctcccag agaccccccg actcttccca ctcaaaatcc
     2401 gttgccgggc tgacctggtc agattgcccc tcaggatgtc ggagcccctg cagagtgtgg
     2461 tggaccacat ggccacccac cttggggtgt ccccaagcag gatccttttg ctttttggag
     2521 agacagagct atcacctact gccactccca ggaccctaaa gctcggagtg gctgacatca
     2581 ttgactgtgt ggtactaaca agttctccag aggccacaga gacgtcccaa cagctccagc
     2641 tccgggtgca gggaaaggag aaacaccaga cactggaagt ctcactgtct cgagattccc
     2701 ctctaaagac cctcatgtcc cactatgagg aggccatggg actgtcggga cggaagctct
     2761 ccttcttctt tgatgggaca aagctttcag gcagggagct gccagctgac ctgggcatgg
     2821 aatctgggga cctcattgag gtctggggct gaGATatcca tcacactggc ggccgctcga
     2881 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     2941 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3001 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3061 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3121 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3181 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3241 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3301 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3361 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3421 tacctagaag ttcctattcc gaagttccta ttctctagaa agtataggaa cttccttggc
     3481 caaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3541 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     3601 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     3661 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg
     3721 ggatttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     3781 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc
     3841 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     3901 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     3961 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4021 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4081 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4141 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4201 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg
     4261 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4321 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4381 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg
     4441 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4501 atagcacgta ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     4561 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     4621 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4681 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4741 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     4801 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4861 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4921 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4981 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     5041 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     5101 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5161 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5221 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5281 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5341 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5401 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5461 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5521 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5581 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5641 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5701 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5761 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5821 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5881 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5941 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6001 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6061 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6121 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6181 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6241 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6301 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6361 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6421 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6481 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6541 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6601 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6661 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6721 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6781 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6841 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6901 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6961 c
//

