LOCUS       pNWA194_(pCMV6-A        8093 bp DNA     circular SYN 24-NOV-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8093)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 24, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     misc_feature    922..1057
                     /label=Synthetic fragment
                     /note="Cloned into pNWA61"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1023..1049
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1068..2327
                     /codon_start=1
                     /label=WT POLI CDS aa 1-420
                     /translation="MELADVGAAASSQGVHDQVLPTPNASSRVIVHVDLDCFYAQVEMI
                     SNPELKDKPLGVQQKYLVVTCNYEARKLGVKKLMNVRDAKEKCPQLVLVNGEDLTRYRE
                     MSYKVTELLEEFSPVVERLGFDENFVDLTEMVEKRLQQLQSDELSAVTVSGHVYNNQSI
                     NLLDVLHIRLLVGSQIAAEMREAMYNQLGLTGCAGVASNKLLAKLVSGVFKPNQQTVLL
                     PESCQHLIHSLNHIKEIPGIGYKTAKCLEALGINSVRDLQTFSPKILEKELGISVAQRI
                     QKLSFGEDNSPVILSGPPQSFSEEDSFKKCSSEVEAKNKIEELLASLLNRVCQDGRKPH
                     TVRLIIRRYSSEKHYGRESRQCPIPSHVIQKLGTGNYDVMTPMVDILMKLFRNMVNVKM
                     PFHLTLLSVCFCNLKALNTAK"
     misc_feature    2352..3239
                     /label=WT POLI 421 - 715
     primer_bind     complement(3301..3317)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3328..3950
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3979..4308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4159..4294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4375..5169
                     /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    5343..5464
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5513..5529)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5537..5553
                     /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(5561..5591)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5606..5627
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5915..6500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6671..7531)
                     /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(7532..7636)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7663..8091
                     /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 tgtacccata cgatgttcca gattacgctg cgatcgccga tatccatatg gagctggccg
     1081 acgtgggagc cgctgccagc tcccaggggg tgcacgatca ggtgctgcca actcccaacg
     1141 cttctagtag agtgatcgtg catgtggacc tggattgctt ttacgcccag gtggagatga
     1201 ttagcaatcc tgaactgaag gacaaaccac tgggcgtgca gcagaaatac ctggtggtga
     1261 catgcaacta tgaagctcgc aagctggggg tgaagaaact gatgaatgtg cgggacgcca
     1321 aggagaaatg tccccagctg gtgctggtga acggggagga tctgacccgg tacagagaaa
     1381 tgtcttataa ggtgacagag ctgctggagg agttcagtcc tgtggtggag cgcctgggct
     1441 tcgacgaaaa ttttgtggat ctgacagaga tggtggaaaa gcggctgcag cagctgcagt
     1501 ctgacgagct gagtgccgtg actgtgagtg gacacgtgta caacaatcag tcaattaacc
     1561 tgctggatgt gctgcatatc agactgctgg tgggcagcca gattgctgcc gagatgaggg
     1621 aagctatgta taaccagctg ggactgaccg gatgtgctgg agtggcctca aataagctgc
     1681 tggccaaact ggtcagcggc gtgtttaaac ccaaccagca gacagtgctg ctgcctgagt
     1741 cttgccagca cctgatccat agtctgaatc acatcaagga aattcccggc atcgggtaca
     1801 agactgctaa atgtctggag gccctgggaa tcaacagcgt gcgcgacctg cagacctttt
     1861 cccctaagat tctggagaaa gaactgggca tttccgtggc ccagagaatc cagaagctgt
     1921 ctttcggaga ggacaattcc ccagtgatcc tgtctggccc ccctcagagc ttctccgagg
     1981 aagatagctt taagaaatgc tcaagcgagg tggaagctaa gaacaaaatt gaggaactgc
     2041 tggcctccct gctgaatagg gtgtgtcagg atgggcgcaa acctcacacc gtgcggctga
     2101 tcattcggag atactcctct gagaagcact atggaaggga atcacgccag tgccctattc
     2161 caagccatgt gatccagaag ctgggaacag gcaactatga cgtgatgact cctatggtgg
     2221 acatcctgat gaaactgttc cgcaacatgg tgaatgtgaa gatgcctttc catctgaccc
     2281 tgctgtctgt gtgcttttgt aacctgaaag ctctgaatac agccaagtaa tctcgagtgg
     2341 atatccatat gaaagggctg attgactact atctgatgcc atctctgagt accacatcac
     2401 ggagcggaaa gcactccttc aagatgaaag acactcacat ggaagacttt cctaaggata
     2461 aagagaccaa tagagatttc ctgccaagtg gcaggatcga gtcaacccgg acaagagaat
     2521 ctcccctgga cactaccaac ttcagtaagg agaaggatat taacgaattt ccactgtgca
     2581 gcctgcctga gggagtggac caggaagtgt ttaagcagct gcctgtggat attcaggagg
     2641 aaatcctgag tgggaagtca agagagaaat tccaggggaa gggcagcgtg tcttgtccac
     2701 tgcacgcttc caggggcgtg ctgagcttct tttccaagaa acagatgcag gacatcccca
     2761 ttaaccctag agatcatctg agttcaagca aacaggtgtc ctctgtgtcc ccatgtgagc
     2821 ccggcacctc tgggtttaac agttcaagct cctcttacat gagttcacag aaggactatt
     2881 cctactatct ggacaatcgg ctgaaagatg aaagaatctc tcagggacct aaggagccac
     2941 agggcttcca ctttacaaac agcaatccag ctgtgtcagc cttccatagc tttcccaacc
     3001 tgcagagcga gcagctgttc tccagaaatc acacaactga cagtcataaa cagactgtgg
     3061 ccaccgattc acacgaaggc ctgacagaga atagggaacc agacagcgtg gatgagaaga
     3121 ttactttccc atccgacatc gatccccagg tgttttacga gctgcccgaa gctgtgcaga
     3181 aagagctgct ggccgaatgg aagaggaccg ggtccgattt ccacatcgga cataagtaag
     3241 gatccaagct tctcgagacg cgttaagcgg ccgcactcga ggtttaaacg gccggccgcg
     3301 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     3361 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     3421 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     3481 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     3541 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3601 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3661 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3721 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3781 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3841 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3901 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3961 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4021 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4081 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4141 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4201 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4261 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4321 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4381 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4441 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4501 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4561 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4621 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4681 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4741 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4801 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4861 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4921 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4981 ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca ggacatagcg
     5041 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5101 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5161 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5221 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5281 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5341 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5401 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5461 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5521 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5581 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5641 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5701 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5761 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5821 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5881 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5941 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6001 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6061 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6121 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6181 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6241 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6301 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6361 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6421 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     7681 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7741 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7801 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7861 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7921 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7981 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     8041 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat ttt
//
