LOCUS       Exported                8090 bp ds-DNA     circular SYN 26-JUL-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8090)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 26, 2019 from SnapGene Viewer 4.3.10
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8090
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1014..1023
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1023..1046
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1065..2834
                     /codon_start=1
                     /label=WT POLI CDS aa 1 - 590
                     /translation="MELADVGAAASSQGVHDQVLPTPNASSRVIVHVDLDCFYAQVEMI
                     SNPELKDKPLGVQQKYLVVTCNYEARKLGVKKLMNVRDAKEKCPQLVLVNGEDLTRYRE
                     MSYKVTELLEEFSPVVERLGFDENFVDLTEMVEKRLQQLQSDELSAVTVSGHVYNNQSI
                     NLLDVLHIRLLVGSQIAAEMREAMYNQLGLTGCAGVASNKLLAKLVSGVFKPNQQTVLL
                     PESCQHLIHSLNHIKEIPGIGYKTAKCLEALGINSVRDLQTFSPKILEKELGISVAQRI
                     QKLSFGEDNSPVILSGPPQSFSEEDSFKKCSSEVEAKNKIEELLASLLNRVCQDGRKPH
                     TVRLIIRRYSSEKHYGRESRQCPIPSHVIQKLGTGNYDVMTPMVDILMKLFRNMVNVKM
                     PFHLTLLSVCFCNLKALNTAKKGLIDYYLMPSLSTTSRSGKHSFKMKDTHMEDFPKDKE
                     TNRDFLPSGRIESTRTRESPLDTTNFSKEKDINEFPLCSLPEGVDQEVFKQLPVDIQEE
                     ILSGKSREKFQGKGSVSCPLHASRGVLSFFSKKQMQDIPINPRDHLSSSKQVSSVSPCE
                     PGTSGFNSSSSSYM"
     misc_feature    2835..2858
                     /label=Insert
     misc_feature    2859..3236
                     /label=WT POLI aa 591 - 715
     primer_bind     complement(3298..3314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3325..3947
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3976..4305
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4372..5166
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5340..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5510..5526)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5534..5550
                     /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(5558..5588)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5603..5624
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5912..6497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6668..7528)
                     /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(7529..7633)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7660..8090,1..25)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccacca
     1021 tggattacaa ggatgacgac gataaggcga tcgccgatat ccatatggag ctggccgacg
     1081 tgggagccgc tgccagctcc cagggggtgc acgatcaggt gctgccaact cccaacgctt
     1141 ctagtagagt gatcgtgcat gtggacctgg attgctttta cgcccaggtg gagatgatta
     1201 gcaatcctga actgaaggac aaaccactgg gcgtgcagca gaaatacctg gtggtgacat
     1261 gcaactatga agctcgcaag ctgggggtga agaaactgat gaatgtgcgg gacgccaagg
     1321 agaaatgtcc ccagctggtg ctggtgaacg gggaggatct gacccggtac agagaaatgt
     1381 cttataaggt gacagagctg ctggaggagt tcagtcctgt ggtggagcgc ctgggcttcg
     1441 acgaaaattt tgtggatctg acagagatgg tggaaaagcg gctgcagcag ctgcagtctg
     1501 acgagctgag tgccgtgact gtgagtggac acgtgtacaa caatcagtca attaacctgc
     1561 tggatgtgct gcatatcaga ctgctggtgg gcagccagat tgctgccgag atgagggaag
     1621 ctatgtataa ccagctggga ctgaccggat gtgctggagt ggcctcaaat aagctgctgg
     1681 ccaaactggt cagcggcgtg tttaaaccca accagcagac agtgctgctg cctgagtctt
     1741 gccagcacct gatccatagt ctgaatcaca tcaaggaaat tcccggcatc gggtacaaga
     1801 ctgctaaatg tctggaggcc ctgggaatca acagcgtgcg cgacctgcag accttttccc
     1861 ctaagattct ggagaaagaa ctgggcattt ccgtggccca gagaatccag aagctgtctt
     1921 tcggagagga caattcccca gtgatcctgt ctggcccccc tcagagcttc tccgaggaag
     1981 atagctttaa gaaatgctca agcgaggtgg aagctaagaa caaaattgag gaactgctgg
     2041 cctccctgct gaatagggtg tgtcaggatg ggcgcaaacc tcacaccgtg cggctgatca
     2101 ttcggagata ctcctctgag aagcactatg gaagggaatc acgccagtgc cctattccaa
     2161 gccatgtgat ccagaagctg ggaacaggca actatgacgt gatgactcct atggtggaca
     2221 tcctgatgaa actgttccgc aacatggtga atgtgaagat gcctttccat ctgaccctgc
     2281 tgtctgtgtg cttttgtaac ctgaaagctc tgaatacagc caagaaaggg ctgattgact
     2341 actatctgat gccatctctg agtaccacat cacggagcgg aaagcactcc ttcaagatga
     2401 aagacactca catggaagac tttcctaagg ataaagagac caatagagat ttcctgccaa
     2461 gtggcaggat cgagtcaacc cggacaagag aatctcccct ggacactacc aacttcagta
     2521 aggagaagga tattaacgaa tttccactgt gcagcctgcc tgagggagtg gaccaggaag
     2581 tgtttaagca gctgcctgtg gatattcagg aggaaatcct gagtgggaag tcaagagaga
     2641 aattccaggg gaagggcagc gtgtcttgtc cactgcacgc ttccaggggc gtgctgagct
     2701 tcttttccaa gaaacagatg caggacatcc ccattaaccc tagagatcat ctgagttcaa
     2761 gcaaacaggt gtcctctgtg tccccatgtg agcccggcac ctctgggttt aacagttcaa
     2821 gctcctctta catgtaatct cgagtggata tccatatgag ttcacagaag gactattcct
     2881 actatctgga caatcggctg aaagatgaaa gaatctctca gggacctaag gagccacagg
     2941 gcttccactt tacaaacagc aatccagctg tgtcagcctt ccatagcttt cccaacctgc
     3001 agagcgagca gctgttctcc agaaatcaca caactgacag tcataaacag actgtggcca
     3061 ccgattcaca cgaaggcctg acagagaata gggaaccaga cagcgtggat gagaagatta
     3121 ctttcccatc cgacatcgat ccccaggtgt tttacgagct gcccgaagct gtgcagaaag
     3181 agctgctggc cgaatggaag aggaccgggt ccgatttcca catcggacat aagtaaggat
     3241 ccaagcttct cgagacgcgt taagcggccg cactcgaggt ttaaacggcc ggccgcggtc
     3301 atagctgttt cctgaacaga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     3361 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3421 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3481 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3541 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3601 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3661 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3721 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3781 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     3841 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     3901 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     3961 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4021 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4081 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4141 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4201 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4261 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4321 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4381 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4441 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4501 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4561 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4621 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4681 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4741 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4801 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4861 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4921 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4981 tctggattca tcgactgtgg ccggctgggt gtggccgacc gctatcagga catagcgttg
     5041 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5101 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5161 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5221 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5281 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5341 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5401 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5461 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     5521 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     5581 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5641 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5701 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5761 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5821 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6421 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6481 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6541 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6601 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6661 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6721 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6781 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6841 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6901 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6961 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7021 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7081 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7141 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7201 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7261 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7321 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7381 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7441 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7501 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7561 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7621 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat
     7681 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     7741 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7801 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7861 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7921 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7981 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     8041 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
//
