LOCUS       Exported                8201 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 8201)
  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..8201
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV forward primer
     protein_bind    820..859
                     /label=TetR binding sites
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    967..988
                     /label=MCS
     misc_feature    1002..1037
                     /label=VSV
     misc_feature    1038..4163
                     /label=CenpB-INCENP
                     /note="/vntifkey=21"
     misc_feature    4182..4200
                     /label=MCS
     primer_bind     4212..4229
                     /label=BGH reverse primer
     polyA_site      4218..4442
                     /label=BGH pA
     rep_origin      4488..4916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4930..5259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5110..5245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5307..5354
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5373..5747
                     /codon_start=1
                     /label=Zeo(R)
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5877..5998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6047..6063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6071..6087
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(6095..6125)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6140..6161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6449..7034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7205..8065)
                     /codon_start=1
                     /label=Amp(R)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8066..8170)
                     /gene="bla"
                     /label=AmpR promoter
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 ttttggaacc
      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 cttggtaccg gatccgccac catgtacacc gacatcgaga
     1021 tgaaccggtt gggcaaggaa ttcggcccca agaggcgaca gctgacgttc cgggagaagt
     1081 cacggatcat ccaggaggtg gaggagaatc cggacctgcg caagggcgag atcgcgcggc
     1141 gcttcaacat cccgccgtcc acgctgagca cgatcctgaa gaacaagcgc gccatcctgg
     1201 cgtcggagcg caagtacggg gtggcctcca cctgccgcaa gaccaacaag ctgtctccct
     1261 acgacaagct cgagggcttg ctcatcgcct ggttccagca gatccgcgcc gccggcctgc
     1321 cggtcaaggg catcatcctc aaggagaagg cgctgcgcat agccgaggag ctgggcatgg
     1381 acgacttcac cgcctccaac ggctggctgg accgcttccg ccggcgccac ggcgtggtgt
     1441 cctgcagcgg cgtggcccgc gcccgcgcgc gaaacgctgc cccccgcacc ccggcggcgc
     1501 ctgccagtcc ggccgcggtg ccctcggagg gcgggagcac tgaattcagc aaagagccag
     1561 agctgatgcc caaaacacct tctcagaaga accgacggaa gaagagacgg atttcttatg
     1621 ttcaggatga aaacagagat cccatcagga gaaggttatc ccgcagaaag tctcggagca
     1681 gccagcagag cacccgacgc ctccgcagca aggacagtgt agagaagctg gctacagtgg
     1741 tcggggagaa cggctccgtt ctgcggcgtg tgacccgtgc tgcggctgca gctgccgcgg
     1801 ctaccatggc attggctgca ccttcttcac ccacccctga gtctcccacg atgctgacta
     1861 agaagcccga ggataaccac acccagtgcc agctggtgcc tgtggtggag atcggcatca
     1921 gtgagcgcca gaatgctgag cagcatgtca cccagctcat gtccaccgag cctctgcccc
     1981 gcactctgtc cccgactcca gcttcagcca cagctccaac ctcccagggc atcccgacat
     2041 cagatgagga atcaacacct aagaagtcga aggccaggat actggagtcc atcacagtga
     2101 gctccctgat ggctacaccc caggacccca agggtcaagg ggtcgggacg gggcggtctg
     2161 cgtctaagct caggattgcg caggtctccc ctggcccacg ggactcgcca gcctttccag
     2221 attctccatg gcgggagcgg gtgctggctc ccatcctgcc ggataacttc tccacgccca
     2281 cgggctctcg cacggactct caatcggtgc ggcacagccc gatcgccccg tcttccccga
     2341 gtccccgagt cttagcccag aagtactctc tggtggccaa acaggaaagt gttgtccgca
     2401 gggcgagcag aaggcttgcc aagaagactg ccgaagagcc agctgcctct ggccgcatca
     2461 tctgtcacag ttacctggag aggctcctga atgttgaggt gccccagaaa gttggttctg
     2521 agcagaagga accccccgag gaggctgagc ctgtggcggc agctgagcca gaggtccctg
     2581 agaacaacgg aaataactcg tggccccaca atgacacaga gatcgccaac agcacaccca
     2641 acccgaagcc tgcagccagc agcccggaaa caccctctgc agggcagcaa gaggccaaga
     2701 cggaccaagc agatggaccc agagagccac cgcagagtgc caggaggaag cgcagctaca
     2761 agcaggccgt gagtgagctg gacgaggagc agcacctgga ggatgaggag ctgcagcccc
     2821 ccaggagcaa gaccccttcc tcaccctgcc cagccagcaa ggtggtacgg cccctccgga
     2881 cctttctgca cacagtgcag aggaaccaga tgctcacgac cccgacctca gccccacgca
     2941 gcgtcatgaa gtcctttatt aagcgcaaca ctcccctgcg catggacccc aagtgcagct
     3001 tcgtcgagaa ggagcggcag cgcctggaga atctgcggcg gaaggaggag gccgagcagc
     3061 tgcgcaggca gaaggtggag gaggacaagc ggcggcggct ggaggaggtg aagctgaagc
     3121 gtgaggaacg cctccgcaag gtgctgcagg cccgcgagcg ggtggagcag atgaaggagg
     3181 agaagaagaa gcagattgag cagaagtttg ctcagatcga cgagaagact gagaaggcca
     3241 aggaggagcg gctggcagag gagaaggcca agaaaaaggc ggcggccaag aagatggagg
     3301 aggtggaagc acgcaggaag caggaagagg atgcacgtag gctcaggtgg ctgcagcagg
     3361 aggaggaaga gcggcggcac caagagctgc tgcagaagaa gaaggaagag gagcaggagc
     3421 ggctgcggaa ggcggccgag gctaagcggc tggcagagca gcgggagcag gagcggcggg
     3481 agcaggagcg gcgcgagcag gagcggcgcg agcaggagcg gcgggagcag gagcggcgcg
     3541 agcaggagcg acagctggca gagcaggagc gtcggcggga gcaggagcgg ctccaggccg
     3601 agagggagct gcaggagcgg gagaaggccc tgcggctgca gaaggagcag ctgcagaggg
     3661 aactggagga gaagaagaag aaggaagagc agcagcgtct ggctgagcgg cagctgcagg
     3721 aggagcaaga gaagaaagcc aaggaggcag taggggccag caaggccctg aatgtgactg
     3781 tggacgtgca gtctccagct tgtacctcat atcaaatgac tccgcaaggg cacagggccc
     3841 ctcccaagat caacccagat aactacggga tggatctgaa tagcgacgac tccaccgatg
     3901 atgaggccca tccccggaag cccatcccca cctgggcccg aggcaccccg ctcagccagg
     3961 ctatcattca ccagtactac cacccaccga accttctgga gctctttgga accattctcc
     4021 cactggactt ggaggatatc ttcaagaaga gcaagccccg ctatcacaag cgcaccagct
     4081 ctgctgtctg gaactcaccg cccctgcagg gcgccagggt ccccagcagc ctggcctaca
     4141 gcctgaagaa gcacaagctt tgactcgagt agatgactag tctagagggc ccgtttaaac
     4201 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4261 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4321 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4381 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     4441 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     4501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5161 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     5221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     5281 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     5341 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     5401 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     5461 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     5521 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     5581 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     5641 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     5701 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     5761 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5821 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5881 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5941 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6001 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6061 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6121 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6181 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6241 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6301 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6361 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6961 gcaaacaaac caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8161 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
