LOCUS       Exported                7859 bp ds-DNA     circular SYN 28-AUG-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pNOC-ARS-stacked-NeoR-GeNL-Ca.dna
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7859)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 31, 2017 from SnapGene Viewer 4.0.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7859)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      5..781)
                     /label=HS terminator
     CDS             complement(797..1588))
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        1601..2781)
                     /label=Ribi
     CDS             2821..3498)
                     /codon_start=1
                     /label=NeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFT"
     CDS             3505..3687)
                     /codon_start=1
                     /label=Nlux
                     /translation="EDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLSG
                     ENGLKIDIHVIIPYEG"
     CDS             3688..4227)
                     /codon_start=1
                     /label=calcium binding domain
                     /translation="PWMHDQLTEEQIAEFKEAFSLFDKDGDGTITTRELGTVMRSLGQN
                     PTEAELQDMINEVDADGNGTIYFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIS
                     AAQLRHVMTNLGVKLTDEEVDEMIREADIDGDGQVNYEEFEQMMTAKGGSKRRWKKNFI
                     AVSAANRFKKISSSGAL"
     CDS             4150..4227)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             4234..4551)
                     /codon_start=1
                     /label=Nlux
                     /translation="LSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTPNMIDY
                     FGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCERIL
                     AG"
     CDS             4552..4578)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      4616..5499)
                     /label=LDSP terminator
     rep_origin      complement(5573..6161))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6332..7192))
                     /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"
     promoter        complement(7193..7297))
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7334..7837)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
ORIGIN
        1 taatggatgg gaagtcggta agtgaaaagg gctttgtgta aggtggatgt caaattttgc
       61 caacccgtgc ctctgtgttc gcagtgtagc aaggaagccc acaccaccgc ctcatattca
      121 aatcgaaatc atcgaagaaa tcggcgagga tctcatgccc caaagctgtc atcaaatgtg
      181 tcccaccaca cgaagatgcc gagtaaatga cgtgagttgt cctatgattg gcaggatgtg
      241 gcatttaaag tccaaaaggc cctgcttggt cggtcacctt ttgctcgcct ccccctcccg
      301 tcacacccta aacacgaatg cagaagaaaa tcatgtgggg aagcaacgat ttagaaatgg
      361 gaatggaggc tcttcacttc ctgaatctga ccaagctggc ggacacaacc gtagagggat
      421 ctttgacctg catcaataca ccgacatgtt aataaatcga gtaccaaaga aaagctactg
      481 aagtcagttt cggattgtgt tgtagaatct tttgagtcgt cactggaatg ctatgctcgc
      541 ttcaatcgtc gattagcttt gtaaattatt tctggtggca aggcggggtt caagtagcaa
      601 gtgggggcat ggcagcaccc ctccgagcct cgttcgttca ggtgtttttg ggacctttcc
      661 ctcccttcct gtcttcagac gcttttattt tcctttccgt tttctactag tgacaaaaaa
      721 agggcacaca ctctttactc gctcacaccc tcgtactcgg acacgcgctg ttaccgactc
      781 tggtacctca agcgctgaag aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat
      841 cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg ccaagctctt
      901 cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca cccagccggc
      961 cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc aagcaggcat
     1021 cgccatgggt cacgacgaga tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca
     1081 gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg acaagaccgg
     1141 cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg
     1201 tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat actttctcgg
     1261 caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat agcagccagt
     1321 cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc gtcgtggcca
     1381 gccacgatag ccgcgctgcc tcgtcttgca gttcattcag ggcaccggac aggtcggtct
     1441 tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca tcagagcagc
     1501 cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg gccggagaac
     1561 ctgcgtgcaa tccatcttgt tcaatcatgg gcccctcgag ggttgcgtgt gtatctgtgt
     1621 gcagtggata ttgttaccga gtttggtgag cgtgagtccg ttagtgccct ggtggtggtg
     1681 gattaggaga gtgggtgact cggtgtccat ggctttcttc gctcattata ggaggggaaa
     1741 ggaatgaggg agggtgggga gaccgcgtct gttgttgacc accgatttac ttcttgcctc
     1801 ccttcccctc cctcccctca atccgtacga cacaaatagt agccgagtgt ctgctgcaga
     1861 gcgcatgatt agtgtggtag acaacgaggg agggaaggat gtacagggca tggcacggag
     1921 aagcgatggt ggccaggaag aggagaggtc gtgagaacag gatgtgttgc gaatggataa
     1981 aaacagaaag cgatggctct gggcttcgaa agcaggggac attaggacgt gtagaccatc
     2041 tcgacggatc cctctgtatc tctgttgtgc gtgaatgttt tctgtgcacg tgtagtgtgt
     2101 gagagtagaa cccgggaact cgaacagaga aaagcatggg tggctgtggt gtggaggctt
     2161 cgttcccacc acatgccctt ctccttcgcc tcgcctctcc ctgccttctt ccacgcaccc
     2221 ttgcgcccct cgttctcaat acctggctca cttccaccat tcaaacaacc atcacgatac
     2281 aggcatttat ctatcgttga agacttcttc ctccggtaga tcttagccaa ggtaagaaga
     2341 ggggcatgca gcaaggagaa agaaatgatg catgatgagg aatagaaggg gaggagggag
     2401 ggatatgatg ggaagcgaaa gcgcatattc tggtggtctg ctgcctgatg gggagtctag
     2461 ctgtgacact gaggacggtg gctgctggtg gctgcgggcg ctgccttggt gatcaatggg
     2521 agtaaaggga gggaaggagg tagcgtgaac ggatgacgcg gagaagttta ggggtctctt
     2581 tacgtatcgc ccctgccgcc cgcctctctg cgataaatgt gcctgttacc ctgcagcctc
     2641 tattcttcac tgtgttcctg ttttccaaca gcctctattc ttccctgtct tttgttgcag
     2701 tggcgtcatc ctctctttgc cccagtcgtc gttctctcga ctcactcact ccccccctcc
     2761 ttccctccct ccatccacag aatcgattgg cgcgccctcg ttaaccaatt ggggagcggc
     2821 atggtctcta agggggaaga agacaatatg gcctcgctgc ccgctacgca tgaactccac
     2881 atttttggtt ccatcaacgg tgtggatttc gatatggttg gccaagggac cggcaaccca
     2941 aatgacggct atgaagagtt aaatctcaag agcacgaaag gggatctaca gtttagtccg
     3001 tggatacttg tgcctcacat cggatacgga tttcaccaat acctgcccta ccctgacgga
     3061 atgagccctt tccaggccgc tatggttgat ggaagtggtt accaggtcca tcggacgatg
     3121 caatttgagg atggggcgag cctgacggtg aattacagat acacctatga ggggagccat
     3181 ataaagggcg aagcccaagt caaggggacc gggtttccag cggacggacc agtgatgaca
     3241 aatagtttga ctgccgcgga ttggtgtcga tccaaaaaga catatcctaa cgacaagacg
     3301 ataatcagta cgttcaaatg gtcctacacg acgggtaatg gtaaacgata taggagcacg
     3361 gcgcggacta cctatacctt cgcaaaaccg atggctgcaa attacttgaa gaaccagcct
     3421 atgtatgtct tccgcaagac cgagctgaag cacagcaaga cggaactcaa ttttaaggaa
     3481 tggcagaaag cctttacggg gttcgaggac ttcgtgggcg actggcggca gacggctggc
     3541 tataacctcg accaggtgct ggagcagggc ggtgtgtcct cgcttttcca gaacctgggc
     3601 gtctccgtga ccccaatcca gcgcatcgtc ctgagcggcg agaacggcct gaagatcgac
     3661 atccacgtca tcattcccta cgaggggcct tggatgcacg atcagctcac tgaggaacaa
     3721 attgcagagt tcaaggaagc gttctcactt tttgataagg atggggatgg gactataacg
     3781 accagagagc taggaaccgt aatgcgttcg ctgggccaaa accctactga ggcggagttg
     3841 caagatatga ttaacgaggt ggacgctgat ggcaatggaa cgatttattt tcccgagttt
     3901 ttgacgatga tggcccggaa gatgaaggat accgactcag aggaggagat ccgtgaggca
     3961 tttcgggtgt tcgacaaaga cggaaacgga tacatctccg ctgcccagct cagacacgtc
     4021 atgacgaact tgggcgtgaa actgacggac gaggaggtcg atgagatgat tcgggaagcg
     4081 gacatagacg gtgatgggca ggtgaactac gaggagtttg agcagatgat gactgccaag
     4141 ggggggtcga agcgccgttg gaagaagaac ttcattgcgg tctccgcggc caaccgattc
     4201 aagaaaatca gttcttccgg cgcgcttgag ttgctgagcg gggaccaaat gggccagatc
     4261 gagaagatct tcaaggttgt gtaccccgtg gacgatcacc acttcaaggt gatactgcac
     4321 tacgggacgc tggtcatcga cggtgtgacg ccgaacatga ttgactactt cggccgtcct
     4381 tacgagggca ttgccgtgtt cgacggcaag aagatcacgg tgaccggcac cctgtggaac
     4441 ggcaataaga tcattgacga gcggctgatc aacccggatg gctcgctgtt gttccgcgtg
     4501 accatcaacg gggtaaccgg ctggcgcctg tgcgagcgca tcctggccgg atacccctac
     4561 gacgtgcccg actacgcggg aagcggcagg cctacgcgtg agtcgcgatg agctcgaaag
     4621 atccaagaga gacgagtaga gatttttttt ttgggattga tgtttgtcgt tctttgagtt
     4681 gtcgtcgagt tacgcctttt gtaagaatgt tccgcaggag aggaggagga tgggcatgag
     4741 tgagggtgag agggcttgcc cgcttttttt tttaaaaacg ctgctcttcg tggttgtcaa
     4801 acaaaccccc catagaaacg attttgttac ggtgcggtcc agacgtcact tgaatggctc
     4861 cgcggaaagg ccagggaggg aaggggggag ggaggaaaca tgaaacatgt tgaacggctc
     4921 aacagggttt gggggacaag agaggtagcg ccctgatgga ctgctccctc ccctcctttc
     4981 cctcaatgtc tcattcatcc atgcttcccc cttctctctc tcccctccgt tccatccccc
     5041 gcgggcgtgg tagtggcgtg atgggatcca ctaaaatgta cgtgtaagaa aagccggtga
     5101 gcttacgctt ttgtgaaagt gggagtacga gtgttgtgtg tgtgtgtagt ggtttcagac
     5161 cccagacaga ggcgaagcag aaaaagcaga cgatgaagac gacgaagaaa tgagcagtct
     5221 atttttatcg tggaaacaga agaggtgata tcgtctcgtt ctttgttatc acctaccccg
     5281 cgtgcatgta catgcagcct ttttattttg taatctttcc cgaaaaatca accgccacct
     5341 cccccccgcc ttctctcacc catcatcttc tcctgtttat cttctacttt acactagatc
     5401 gcatggcaca tctccctcgc aatccatcgg tgcaaccatc atcgatccca ctcctccctc
     5461 cctccctccc tccctcctcg catctcttct aagaaatccg ctagtgttta aacatgtgag
     5521 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5581 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5641 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5701 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5761 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5821 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5881 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5941 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6001 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6061 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6121 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6181 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6241 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7321 ctgggtcctt ttcatcacgt gctataaaaa taattataat ttaaattttt taatataaat
     7381 atataaatta aaaatagaaa gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc
     7441 gaagatgtaa aagactctag ggggatcgcc aacaaatact accttttatc ttgtacttcc
     7501 tgctctcagg tattaatgcc gaattgtttc atcttgtctg tgtagaagac cacacacgaa
     7561 aatcctgtga ttttacattt tacttatcgt taatcgaatg tatatctatt taatctgctt
     7621 ttcttgtcta ataaatatat atgtaaagta cgctttttgt tgaaattttt taaacctttg
     7681 tttatttttt tttcttcatt ccgtaactct tctaccttct ttatttactt tctaaaatcc
     7741 aaatacaaaa cataaaaata aataaacaca gagtaaattc ccaaattatt ccatcattaa
     7801 aagatacgag gcgcgtgtaa gttacaggca agcgatccgt ctaagaaacc ggttaaggc
//
