LOCUS       Exported                6131 bp ds-DNA     circular SYN 20-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6131)
  AUTHORS   UMC-Utrecht
  TITLE     Direct Submission
  JOURNAL   Exported Mar 8, 2018 from SnapGene Viewer 4.1.7
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6131
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        6..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV forward primer
     promoter        634..652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             687..719
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     regulatory      726..735
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    join(732..1190,1193..1478,1480..1766)
                     /label=Aurora B
     misc_feature    1191..1193
                     /label=Leu to Ala
                     /label=Leu to Gly
     misc_feature    1479..1481
                     /label=His to Tyr
     misc_feature    1764..1766
                     /label=STOP
     promoter        complement(1842..1860)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1880..1897
                     /label=BGH reverse primer
     polyA_signal    1886..2110
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2242..2830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    3159..3206
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(3438..4226)
                     /codon_start=1
                     /label=Neo(R)
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(4261..4618)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             complement(4678..5538)
                     /codon_start=1
                     /label=Amp(R)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5539..5643)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5670..6125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga ctatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      601 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      661 agcttggtac cggatccgcc accatgtaca ccgacatcga gatgaaccgg ttgggcaagg
      721 aattcgccac catggcccag aaggagaact cctacccctg gccctacggc cgacagacgg
      781 ctccatctgg cctgagcacc ctgccccagc gagtcctccg gaaagagcct gtcaccccat
      841 ctgcacttgt cctcatgagc cgctccaatg tccagcccac agctgcccct ggccagaagg
      901 tgatggagaa tagcagtggg acacccgaca tcttaacgcg gcacttcaca attgatgact
      961 ttgagattgg gcgtcctctg ggcaaaggca agtttggaaa cgtgtacttg gctcgggaga
     1021 agaaaagcca tttcatcgtg gcgctcaagg tcctcttcaa gtcccagata gagaaggagg
     1081 gcgtggagca tcagctgcgc agagagatcg aaatccaggc ccacctgcac catcccaaca
     1141 tcctgcgtct ctacaactat ttttatgacc ggaggaggat ctacttgatt gcagagtatg
     1201 ccccccgcgg ggagctctac aaggagctgc agaagagctg cacatttgac gagcagcgaa
     1261 cagccacgat catggaggag ttggcagatg ctctaatgta ctgccatggg aagaaggtga
     1321 ttcacagaga cataaagcca gaaaatctgc tcttagggct caagggagag ctgaagattg
     1381 ctgacttcgg ctggtctgtg catgcgccct ccctgaggag gaagacaatg tgtggcaccc
     1441 tggactacct gcccccagag atgattgagg ggcgcatgta caatgagaag gtggatctgt
     1501 ggtgcattgg agtgctttgc tatgagctgc tggtggggaa cccacccttt gagagtgcat
     1561 cacacaacga gacctatcgc cgcatcgtca aggtggacct aaagttcccc gcttctgtgc
     1621 ccacgggagc ccaggacctc atctccaaac tgctcaggca taacccctcg gaacggctgc
     1681 ccctggccca ggtctcagcc cacccttggg tccgggccaa ctctcggagg gtgctgcctc
     1741 cctctgccct tcaatctgtc gcctgacccg ggaatcgaat tctgcagata tccatcacac
     1801 tggcggccgc tcgagtagat gactagtcta gagggcccta ttctatagtg tcacctaaat
     1861 gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1921 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1981 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     2041 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     2101 ggctctatgg cttctgaggc ggaaagaacc agtggcggta atacggttat ccacagaatc
     2161 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2221 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2281 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2341 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2401 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2461 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2521 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2581 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2641 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     2701 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2761 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2821 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2881 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2941 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc
     3001 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg
     3061 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc
     3121 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg
     3181 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt
     3241 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg
     3301 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa
     3361 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc
     3421 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
     3481 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
     3541 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
     3601 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
     3661 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg
     3721 aacagttcgg ctggcgcgag cccctgatgc tcttgatcat cctgatcgac aagaccggct
     3781 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3841 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3901 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3961 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     4021 cacgatagcc gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg
     4081 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     4141 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     4201 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc
     4261 tttgcaaaag cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc
     4321 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg
     4381 ggcggaactg ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc
     4441 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     4501 acacctggtt gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc
     4561 tggggacttt ccacacccta actgacacac attccacagc tggttctttc cgcctcagga
     4621 ctcttccttt ttcaataaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5641 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     5701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6121 aatttacgcg c
//
