LOCUS       Exported                7313 bp ds-DNA     circular SYN 17-AUG-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pNOC-ARS-stacked-NeoR-GeNL.dna
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7313)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 2017 from SnapGene Viewer 4.0.0
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7313)
                     /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..4005)
                     /codon_start=1
                     /label=GeNL
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTGFEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRIVLSGENGLKIDI
                     HVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVTPNMIDYFGRPYE
                     GIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWRLCERILAG"
     CDS             4006..4032)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      4070..4953)
                     /label=LDSP terminator
     rep_origin      complement(5027..5615))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5786..6646))
                     /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(6647..6751))
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6788..7291)
                     /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 cgagggcctg agcggggacc aaatgggcca gatcgagaag
     3721 atcttcaagg ttgtgtaccc cgtggacgat caccacttca aggtgatact gcactacggg
     3781 acgctggtca tcgacggtgt gacgccgaac atgattgact acttcggccg tccttacgag
     3841 ggcattgccg tgttcgacgg caagaagatc acggtgaccg gcaccctgtg gaacggcaat
     3901 aagatcattg acgagcggct gatcaacccg gatggctcgc tgttgttccg cgtgaccatc
     3961 aacggggtaa ccggctggcg cctgtgcgag cgcatcctgg ccggataccc ctacgacgtg
     4021 cccgactacg cgggaagcgg caggcctacg cgtgagtcgc gatgagctcg aaagatccaa
     4081 gagagacgag tagagatttt ttttttggga ttgatgtttg tcgttctttg agttgtcgtc
     4141 gagttacgcc ttttgtaaga atgttccgca ggagaggagg aggatgggca tgagtgaggg
     4201 tgagagggct tgcccgcttt tttttttaaa aacgctgctc ttcgtggttg tcaaacaaac
     4261 cccccataga aacgattttg ttacggtgcg gtccagacgt cacttgaatg gctccgcgga
     4321 aaggccaggg agggaagggg ggagggagga aacatgaaac atgttgaacg gctcaacagg
     4381 gtttggggga caagagaggt agcgccctga tggactgctc cctcccctcc tttccctcaa
     4441 tgtctcattc atccatgctt cccccttctc tctctcccct ccgttccatc ccccgcgggc
     4501 gtggtagtgg cgtgatggga tccactaaaa tgtacgtgta agaaaagccg gtgagcttac
     4561 gcttttgtga aagtgggagt acgagtgttg tgtgtgtgtg tagtggtttc agaccccaga
     4621 cagaggcgaa gcagaaaaag cagacgatga agacgacgaa gaaatgagca gtctattttt
     4681 atcgtggaaa cagaagaggt gatatcgtct cgttctttgt tatcacctac cccgcgtgca
     4741 tgtacatgca gcctttttat tttgtaatct ttcccgaaaa atcaaccgcc acctcccccc
     4801 cgccttctct cacccatcat cttctcctgt ttatcttcta ctttacacta gatcgcatgg
     4861 cacatctccc tcgcaatcca tcggtgcaac catcatcgat cccactcctc cctccctccc
     4921 tccctccctc ctcgcatctc ttctaagaaa tccgctagtg tttaaacatg tgagcaaaag
     4981 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5041 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5101 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5161 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5221 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5281 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5341 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5401 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5461 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5521 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5581 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5641 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5701 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5761 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5821 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5881 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5941 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6001 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6061 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6121 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6181 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6241 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6301 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6361 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6421 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6481 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6541 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6601 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6661 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6721 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgggt
     6781 ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat aaatatataa
     6841 attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt ttccgaagat
     6901 gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgtac ttcctgctct
     6961 caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca cgaaaatcct
     7021 gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct gcttttcttg
     7081 tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc tttgtttatt
     7141 tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa atccaaatac
     7201 aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca ttaaaagata
     7261 cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccggttaa ggc
//
