LOCUS       Exported                8758 bp ds-DNA     circular SYN 07-JUN-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8758)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 7, 2019 from SnapGene Viewer 4.3.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8758
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    716..737
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        752..782
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    790..806
                     /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)."
     primer_bind     814..830
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             1091..1724
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    1771..1896
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2393..2626
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    3152..3269
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3321..3624
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3625..3828
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        3924..3942
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3972..3996
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3996..4406
                     /codon_start=1
                     /label=C20orf24
                     /translation="MSGGRRKEEPPQPQLANGALKVSVWSKVLRSDAAWEDKDEFLDVI
                     YWFRQIIAVVLGVIWGVLPLRGFLGIAGFCLINAGVLYLYFSNYLQIDEEEYGGTWELT
                     KEGFMTSFALFMVCVADSFTTGHLDHLLHCHPL"
     protein_bind    complement(4408..4432)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4464..4487
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4503..4529
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     3'UTR           4533..5038
                     /label=3'UTR-Variant
     misc_feature    4930
                     /label=c.*398G>A
     misc_feature    5059..5610
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5622..6221
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     misc_feature    6238..6826
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6709..6720)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6901..7134
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        7339..7443
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7444..8304
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      join(8475..8758,1..305)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
       61 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      121 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      181 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      241 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      301 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      361 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      421 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      481 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      541 gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
      601 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
      661 ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
      721 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
      781 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
      841 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
      901 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
      961 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
     1021 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
     1081 acttagtagt tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg
     1141 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc
     1201 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta
     1261 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg
     1321 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac
     1381 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag
     1441 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga
     1501 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga
     1561 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     1621 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     1681 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     1741 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     1801 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     1861 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     1921 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1981 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     2041 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     2101 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     2161 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     2221 accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca
     2281 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     2341 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     2401 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     2461 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     2521 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     2581 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     2641 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     2701 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     2761 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     2821 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     2881 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     2941 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     3001 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     3061 gagagagaga cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa
     3121 ttaattcaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     3181 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     3241 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     3301 tttggactag tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3361 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3421 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3481 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3541 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3601 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     3661 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3721 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     3781 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     3841 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccggcgg
     3901 ccgcagatct ctcgaggttg atctaatacg actcactata gggagaccca agctggctag
     3961 ttaagctatc aacaagtttg tacaaaaaag ttggcatgag cggcgggcgg cggaaggagg
     4021 agccgcctca gccgcagctg gccaacgggg ccctcaaagt ctccgtctgg agtaaggtgc
     4081 tgcggagcga cgcggcctgg gaggataagg atgaattttt agatgtgatc tactggttcc
     4141 gacagatcat tgctgtggtc ctgggtgtca tttggggagt tttgccatta cgagggttct
     4201 tgggaatagc aggattctgc ctgatcaatg caggagtcct gtacctctac ttcagcaatt
     4261 acctacagat tgatgaggaa gaatatggtg gcacgtggga gctcacgaag gaagggttta
     4321 tgacctcttt tgccttgttc atggtatgtg tagctgatag ttttacaaca ggtcatttgg
     4381 atcatctttt acactgccat ccattatacc caactttctt gtacaaagtg gttgatctag
     4441 agggcccgcg gttcgaactc gaggactaca aggacgacga tgacaagtcg gccgctggag
     4501 gataccccta cgacgtgccc gactacgcct agtggtgtac agctcccaag tgctccctat
     4561 ccagtccaaa ggaccctctt gattacagca caggaacttg atcgttgggg aaccccagcc
     4621 ccttggaact tggaagaccc gtgtttcctg gaccgcgaat cagtgtgttg ggcatcagtg
     4681 ttttctgcaa gggttgtgac ctgaaacttt ttaaaaacca cccacctttg gggaagcatt
     4741 tctgaattta tccatcacca accatttctt cttggatacc atcaagtaac agctattatt
     4801 tgccaagtgg agctgtcatt taatttgatg cacctctgga ttcagatgaa acattaaatt
     4861 gtcttcctcg attctccatc gggtgtagag tttttaaact atcaatggca tttcaagtct
     4921 tctgaaacaa catggctgta tgtgcgtggt ccatagcaca gtacatgcag catctaataa
     4981 gagtttccat tgtagaatgt tttcacatac ttgaataaat caaatcttta attgagaaaa
     5041 ttccgccccc cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     5101 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     5161 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     5221 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     5281 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     5341 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     5401 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     5461 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat
     5521 gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc acggggacgt
     5581 ggttttcctt tgaaaaacac gatgataata tggccacaac catgaccgag tacaagccca
     5641 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     5701 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     5761 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     5821 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     5881 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     5941 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     6001 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     6061 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     6121 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     6181 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg atgcatcgat ggatcctaat
     6241 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     6301 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     6361 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     6421 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     6481 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     6541 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     6601 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     6661 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6721 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     6781 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctga gatcctttaa
     6841 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     6901 tggaagggct aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc
     6961 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7021 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7081 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta
     7141 gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg
     7201 agaggcccgg gttaattaag gaaagggcta gatcattctt gaagacgaaa gggcctcgtg
     7261 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     7321 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     7381 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     7441 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7501 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7561 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7621 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7681 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7741 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7801 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7861 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7921 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7981 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     8041 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     8101 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     8161 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     8221 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     8281 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     8341 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     8401 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     8461 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8521 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8581 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8641 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8701 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagac
//
