LOCUS       Exported                5198 bp ds-DNA     circular SYN 15-JAN-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5198)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 15 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"
     polyA_signal    2181..2405
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    2689..2736
                     /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             2744..3766
                     /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    3896..4017
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4066..4082)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    4090..4106
                     /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(4114..4144)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    4159..4180
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the
                     presence of cAMP."
     rep_origin      complement(4468..5056)
                     /dnas_title="ori"
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(5227..6087)
                     /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(6088..6192)
                     /dnas_title="AmpR promoter"
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    979..1075
                     /note="3xFLAG insert from annealed oligos"
     CDS             993..1058
                     /note="3xFLAG"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1083..2114
                     /note="NFATC2IP delM delSLD2 aa345 - 419"
                     /translation="AEPVGKRGRWSGGSGAGRGGRGGWGGRGRRPRAQRSPSRGTLDV
                     VSVDLVTDSDEEILEVATARGAADEVEVEPPEPPGPVASRDNSNSDSEGEDRRPAGPP
                     REPVRRRRRLVLDPGEAPLVPVYSGKVKSSLRLIPDDLSLLKLYPPGDEEEAELADSS
                     GLYHEGSPSPGSPWKTKLRTKDKEEKKKTEFLDLDNSPLSPPSPRTKSRTHTRALKKL
                     SEVNKRLQDLRSCLSPKPPQGQEQQGQEDEVVLVEGPTLPETPRLFPLKIRCRADLVR
                     LPLRMSEPLQSVVDHMATHLGVSPSRILLLFGETELSPTATPRTLKLGVADIIDCVVL
                     TSSPEATET"
     source          1..6223
                     /dnas_title="Restriction 1 Restriction Clone"
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 TGGACTACAA GGACCACGAC GGTGACTACA
     1021 AGGACCACGA CATCGACTAC AAAGACGATG ACGATAAAGC CGCCAGTTTA AACGGCGCGC
     1081 CAgcggagcc tgtggggaag cggggccgct ggtccggagg tagcggtgcc ggccgagggg
     1141 gtcggggcgg ctggggcggt cggggccggc gtcctcgggc ccagcggtct ccatcccggg
     1201 gcacgctgga cgtagtgtct gtggacttgg tcaccgacag cgatgaggaa attctggagg
     1261 tcgccaccgc tcgcggtgcc gcggacgagg ttgaggtgga gcccccggag cccccggggc
     1321 cggtcgcgtc ccgggataac agcaacagtg acagcgaagg ggaggacagg cggcccgcag
     1381 gacccccgcg ggagccggtc aggcggcggc ggcggctggt gctggatccg ggggaggcgc
     1441 cgctggttcc ggtgtactcg gggaaggtta aaagcagcct tcgccttatc ccagatgatc
     1501 tatccctcct gaaactctac cctccagggg atgaggaaga ggcagagctg gcagattcga
     1561 gtggtctcta ccatgagggc tccccatcac caggctctcc ctggaagaca aagctgagga
     1621 ctaaggataa agaagagaag aaaaagacag agtttctgga tctggacaac tctcctctgt
     1681 ccccaccttc accaaggacc aaaagcagaa cgcatactcg ggcactcaag aagttaagtg
     1741 aggtgaacaa gcgcctccag gatctccgtt cctgtctgag ccccaagcca cctcagggtc
     1801 aagagcaaca gggccaagag gatgaagtgg tcttggtgga agggcccacc ctcccagaga
     1861 ccccccgact cttcccactc aaaatccgtt gccgggctga cctggtcaga ttgcccctca
     1921 ggatgtcgga gcccctgcag agtgtggtgg accacatggc cacccacctt ggggtgtccc
     1981 caagcaggat ccttttgctt tttggagaga cagagctatc acctactgcc actcccagga
     2041 ccctaaagct cggagtggct gacatcattg actgtgtggt actaacaagt tctccagagg
     2101 ccacagagac gtgaGATatc catcacactg gcggccgctc gagtctagag ggcccgttta
     2161 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2221 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2281 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2341 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     2401 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     2461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt
     2701 ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac
     2761 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     2821 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     2881 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     2941 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggatttca gcgagagcct
     3001 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     3061 actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct
     3121 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     3181 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     3241 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     3301 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     3361 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     3421 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     3481 ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct
     3541 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     3601 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     3661 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     3721 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga
     3781 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3841 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     3901 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3961 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4021 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4201 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4261 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4321 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4381 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4441 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4501 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4561 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4621 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4681 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4741 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4801 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4861 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     4921 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4981 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5041 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5101 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5161 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5221 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5281 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5341 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5401 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5461 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5521 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5581 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5641 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5701 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5761 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5821 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5881 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5941 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6001 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6061 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6121 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6181 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//

