LOCUS       pNWA196_(pCMV6-A        7504 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 7504)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 24, 2023 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7504
                     /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"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    998..2651
                     /label=Synthetic fragment
                     /note="Cloned into pJRM46"
     CDS             1023..1049
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1068..2636
                     /codon_start=1
                     /label=Rev1 321-843
                     /translation="SSTKSTSSVSTFSKAAPSVPSKPSDCNFISNFYSHSRLHHISMWK
                     CELTEFVNTLQRQSNGIFPGREKLKKMKTGRSALVVTDTGDMSVLNSPRHQSCIMHVDM
                     DCFFVSVGIRNRPDLKGKPVAVTSNRGTGRAPLRPGANPQLEWQYYQNKILKGKAADIP
                     DSSLWENPDSAQANGIDSVLSRAEIASCSYEARQLGIKNGMFFGHAKQLCPNLQAVPYD
                     FHAYKEVAQTLYETLASYTHNIEAVSCDEALVDITEILAETKLTPDEFANAVRMEIKDQ
                     TKCAASVGIGSNILLARMATRKAKPDGQYHLKPEEVDDFIRGQLVTNLPGVGHSMESKL
                     ASLGIKTCGDLQYMTMAKLQKEFGPKTGQMLYRFCRGLDDRPVRTEKERKSVSAEINYG
                     IRFTQPKEAEAFLLSLSEEIQRRLEATGMKGKRLTLKIMVRKPGAPVETAKFGGHGICD
                     NIARTVTLDQATDNAKIIGKAMLNMFHTMKLNISDMRGVGIHVNQLVPTNLNPSTCPSR
                     PSVQSS"
     misc_feature    1095..2606
                     /label=Catalytic Core
     primer_bind     complement(2701..2717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2728..3350
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     promoter        3379..3708
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3559..3694
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3775..4569
                     /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    4743..4864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4913..4929)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4937..4953
                     /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(4961..4991)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5006..5027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5315..5900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6071..6931)
                     /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(6932..7036)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7063..7491
                     /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 tatccattct agcaccaagt
     1081 ctacatcctc tgtgagcacc ttttccaagg ccgccccttc tgtgccaagc aagccctccg
     1141 attgcaattt catctctaac ttttatagcc actcccggct gcaccacatc agcatgtgga
     1201 agtgtgagct gaccgagttc gtgaatacac tgcagagaca gtctaacggc atctttccag
     1261 gcagggagaa gctgaagaag atgaagaccg gcagaagcgc cctggtggtg accgacacag
     1321 gcgatatgag cgtgctgaac tcccccaggc accagtcttg catcatgcac gtggacatgg
     1381 attgtttctt tgtgagcgtg ggcatcagga acaggccaga cctgaagggc aagccagtgg
     1441 cagtgacctc caatagggga acaggaagag cacctctgag gccaggagca aacccacagc
     1501 tggagtggca gtactatcag aataagatcc tgaagggcaa ggccgccgac atcccagata
     1561 gctccctgtg ggagaatccc gacagcgccc aggccaacgg catcgattct gtgctgagcc
     1621 gcgccgagat cgcctcttgc agctacgagg cccggcagct gggcatcaag aacggcatgt
     1681 tctttggcca cgccaagcag ctgtgcccta atctgcaggc cgtgccctac gacttccacg
     1741 cctacaagga ggtggcccag accctgtatg agacactggc ctcctacacc cacaacatcg
     1801 aggccgtgtc ttgcgacgag gccctggtgg atatcacaga gatcctggcc gagacaaagc
     1861 tgacacctga cgagtttgcc aacgccgtga gaatggagat caaggatcag acaaagtgtg
     1921 ccgcctctgt gggcatcggc agcaatatcc tgctggcaag gatggcaacc cggaaggcaa
     1981 agcctgacgg acagtatcac ctgaagccag aggaggtgga cgatttcatc aggggacagc
     2041 tggtgaccaa cctgccagga gtgggccact ccatggagtc taagctggcc agcctgggca
     2101 tcaagacctg cggcgatctg cagtatatga caatggccaa gctgcagaag gagttcggcc
     2161 ccaagaccgg ccagatgctg tacaggtttt gtcgcggcct ggacgatcgg cctgtgagaa
     2221 cagagaagga gagaaagtcc gtgtctgccg agatcaatta cggcatcagg ttcacccagc
     2281 caaaggaggc cgaggccttt ctgctgagcc tgtccgagga gatccagcgg agactggagg
     2341 ccaccggcat gaagggcaag agactgacac tgaagatcat ggtgaggaag ccaggagcac
     2401 ctgtggagac agccaagttc ggcggccacg gcatctgcga caacatcgca aggaccgtga
     2461 cactggacca ggcaaccgat aatgccaaga tcatcggcaa ggccatgctg aatatgtttc
     2521 acacaatgaa gctgaacatc tccgatatgc ggggcgtggg catccacgtg aaccagctgg
     2581 tgcctaccaa cctgaatcca tctacatgcc caagcagacc ctccgtgcag tctagctaag
     2641 gatccaagct tctcgagacg cgttaagcgg ccgcactcga ggtttaaacg gccggccgcg
     2701 gtcatagctg tttcctgaac agatcccggg tggcatccct gtgacccctc cccagtgcct
     2761 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     2821 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     2881 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     2941 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     3001 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     3061 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     3121 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     3181 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc
     3241 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt
     3301 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg
     3361 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3421 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3481 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3541 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3601 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3661 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3721 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3781 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3841 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3901 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3961 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4021 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4081 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4141 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4201 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4261 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4321 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4381 ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca ggacatagcg
     4441 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4501 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4561 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4621 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4681 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4741 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4801 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4861 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4921 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4981 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5041 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5101 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5821 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     5881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6121 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6181 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6241 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6301 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6361 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6421 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6481 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6541 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6601 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6661 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6721 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6781 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6841 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6901 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6961 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7021 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     7081 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     7141 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7201 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7261 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7321 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7381 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     7441 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     7501 tatt
//
