LOCUS       Exported                8053 bp ds-DNA     circular SYN 07-FEB-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8053)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 9, 2020 from SnapGene Viewer 5.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8053
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             906..932
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             945..3557
                     /codon_start=1
                     
                     /translation="MTADKEKKRSSSERRKEKSRDAARCRRSKETEVFYELAHELPLPH
                     SVSSHLDKASIMRLAISFLRTHKLLSSVCSENESEAEADQQMDNLYLKALEGFIAVVTQ
                     DGDMIFLSENISKFMGLTQVELTGHSIFDFTHPCDHEEIRENLSLKNGSGFGKKSKDMS
                     TERDFFMRMKCTVTNRGRTVNLKSATWKVLHCTGQVKVYNNCPPHNSLCGYKEPLLSCL
                     IIMCEPIQHPSHMDIPLDSKTFLSRHSMDMKFTYCDDRITELIGYHPEELLGRSAYEFY
                     HALDSENMTKSHQNLCTKGQVVSGQYRMLAKHGGYVWLETQGTVIYNPRNLQPQCIMCV
                     NYVLSEIEKNDVVFSMDQTESLFKPHLMAMNSIFDSSGKGAVSEKSNFLFTKLKEEPEE
                     LAQLAPTPGDAIISLDFGNQNFEESSAYGKAILPPSQPWATELRSHSTQSEAGSLPAFT
                     VPQAAAPGSTTPSATSSSSSCSTPNSPEDYYTSLDNDLKIEVIEKLFAMDTEAKDQCST
                     QTDFNELDLETLAPYIPMDGEDFQLSPICPEERLLAENPQSTPQHCFSAMTNIFQPLAP
                     VAPHSPFLLDKFQQQLESKKTEPEHRPMSSIFFDAGSKASLPPCCGQASTPLSSMGGRS
                     NTQWPPDPPLHFGPTKWAVGDQRTEFLGAAPLGPPVSPPHVSTFKTRSAKGFGARGPDV
                     LSPAMVALSNKLKLKRQLEYEEQAFQDLSGGDPPGGSTSHLMWKRMKNLRGGSCPLMPD
                     KPLSANVPNDKFTQNPMRGLGHPLRHLPLPQPPSAISPGENSKSRFPPQCYATQYQDYS
                     LSSAHKVSGMASRLLGPSFESYLLPELTRYDCEVNVPVLGSSTLLQGGDLLRALDQAT"
     promoter        complement(3608..3626)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3652..3876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3922..4350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4364..4693
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4544..4679
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4760..5554
                     /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    5728..5849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5898..5914)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5922..5938
                     /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(5946..5976)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5991..6012
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6300..6888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7059..7919)
                     /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(7920..8024)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttacca
      901 tggcctaccc ctacgacgtg cccgactacg cctccctcgg atccatgaca gctgacaagg
      961 agaagaaaag gagtagctcg gagaggagga aggagaagtc ccgggatgct gcgcggtgcc
     1021 ggcggagcaa ggagacggag gtgttctatg agctggccca tgagctgcct ctgccccaca
     1081 gtgtgagctc ccatctggac aaggcctcca tcatgcgact ggcaatcagc ttcctgcgaa
     1141 cacacaagct cctctcctca gtttgctctg aaaacgagtc cgaagccgaa gctgaccagc
     1201 agatggacaa cttgtacctg aaagccttgg agggtttcat tgccgtggtg acccaagatg
     1261 gcgacatgat ctttctgtca gaaaacatca gcaagttcat gggacttaca caggtggagc
     1321 taacaggaca tagtatcttt gacttcactc atccctgcga ccatgaggag attcgtgaga
     1381 acctgagtct caaaaatggc tctggttttg ggaaaaaaag caaagacatg tccacagagc
     1441 gggacttctt catgaggatg aagtgcacgg tcaccaacag aggccgtact gtcaacctca
     1501 agtcagccac ctggaaggtc ttgcactgca cgggccaggt gaaagtctac aacaactgcc
     1561 ctcctcacaa tagtctgtgt ggctacaagg agcccctgct gtcctgcctc atcatcatgt
     1621 gtgaaccaat ccagcaccca tcccacatgg acatccccct ggatagcaag accttcctga
     1681 gccgccacag catggacatg aagttcacct actgtgatga cagaatcaca gaactgattg
     1741 gttaccaccc tgaggagctg cttggccgct cagcctatga attctaccat gcgctagact
     1801 ccgagaacat gaccaagagt caccagaact tgtgcaccaa gggtcaggta gtaagtggcc
     1861 agtaccggat gctcgcaaag catgggggct acgtgtggct ggagacccag gggacggtca
     1921 tctacaaccc tcgcaacctg cagccccagt gcatcatgtg tgtcaactac gtcctgagtg
     1981 agattgagaa gaatgacgtg gtgttctcca tggaccagac tgaatccctg ttcaagcccc
     2041 acctgatggc catgaacagc atctttgata gcagtggcaa gggggctgtg tctgagaaga
     2101 gtaacttcct attcaccaag ctaaaggagg agcccgagga gctggcccag ctggctccca
     2161 ccccaggaga cgccatcatc tctctggatt tcgggaatca gaacttcgag gagtcctcag
     2221 cctatggcaa ggccatcctg cccccgagcc agccatgggc cacggagttg aggagccaca
     2281 gcacccagag cgaggctggg agcctgcctg ccttcaccgt gccccaggca gctgccccgg
     2341 gcagcaccac ccccagtgcc accagcagca gcagcagctg ctccacgccc aatagccctg
     2401 aagactatta cacatctttg gataacgacc tgaagattga agtgattgag aagctcttcg
     2461 ccatggacac agaggccaag gaccaatgca gtacccagac ggatttcaat gagctggact
     2521 tggagacact ggcaccctat atccccatgg acggggaaga cttccagcta agccccatct
     2581 gccccgagga gcggctcttg gcggagaacc cacagtccac cccccagcac tgcttcagtg
     2641 ccatgacaaa catcttccag ccactggccc ctgtagcccc gcacagtccc ttcctcctgg
     2701 acaagtttca gcagcagctg gagagcaaga agacagagcc cgagcaccgg cccatgtcct
     2761 ccatcttctt tgatgccgga agcaaagcat ccctgccacc gtgctgtggc caggccagca
     2821 cccctctctc ttccatgggg ggcagatcca atacccagtg gcccccagat ccaccattac
     2881 attttgggcc cacaaagtgg gccgtcgggg atcagcgcac agagttcttg ggagcagcgc
     2941 cgttggggcc ccctgtctct ccaccccatg tctccacctt caagacaagg tctgcaaagg
     3001 gttttggggc tcgaggccca gacgtgctga gtccggccat ggtagccctc tccaacaagc
     3061 tgaagctgaa gcgacagctg gagtatgaag agcaagcctt ccaggacctg agcggggggg
     3121 acccacctgg tggcagcacc tcacatttga tgtggaaacg gatgaagaac ctcaggggtg
     3181 ggagctgccc tttgatgccg gacaagccac tgagcgcaaa tgtacccaat gataagttca
     3241 cccaaaaccc catgaggggc ctgggccatc ccctgagaca tctgccgctg ccacagcctc
     3301 catctgccat cagtcccggg gagaacagca agagcaggtt ccccccacag tgctacgcca
     3361 cccagtacca ggactacagc ctgtcgtcag cccacaaggt gtcaggcatg gcaagccggc
     3421 tgctcgggcc ctcatttgag tcctacctgc tgcccgaact gaccagatat gactgtgagg
     3481 tgaacgtgcc cgtgctggga agctccacgc tcctgcaagg aggggacctc ctcagagccc
     3541 tggaccaggc cacctgagat atccatcaca ctggcggccg ctcgagcatg catctagagg
     3601 gccctattct atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3901 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3961 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4021 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4081 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4141 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4201 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4261 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4321 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4441 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4501 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4561 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4621 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4681 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     4741 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     4801 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4861 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4921 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4981 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5041 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5101 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5161 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5221 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5281 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5341 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5401 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5461 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5521 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5581 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5641 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     5701 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     5761 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5821 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5881 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5941 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6001 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6061 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6121 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6181 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6241 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6301 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6361 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6421 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6481 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6541 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6601 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6661 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6721 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6781 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6841 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6901 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6961 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7021 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7081 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7141 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7201 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7261 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7321 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7381 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7441 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7501 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7561 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7621 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7681 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     7741 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     7801 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7861 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7921 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7981 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8041 gtgccacctg acg
//
