LOCUS       Exported                8357 bp ds-DNA     circular SYN 16-SEP-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8357)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 16, 2019 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8357
                     /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..4511
                     /codon_start=1
                     /label=NENF
                     /translation="MVGPAPRRRLRPLAALALVLALAPGLPTARAGQTPRPAERGPPVR
                     LFTEEELARYGGEEEDQPIYLAVKGVVFDVTSGKEFYGRGAPYNALTGKDSTRGVAKMS
                     LDPADLTHDTTGLTAKELEALDEVFTKVYKAKYPIVGYTARRILNEDGSPNLDFKPEDQ
                     PHFDIKDEF"
     primer_bind     complement(4209..4252)
                     /label=NENF-inner_R
     primer_bind     4227..4264
                     /label=NENF-inner_F
     misc_feature    4236..4247
                     /label=Synonymous mutations to silence sgRNA seed region
     protein_bind    complement(4515..4537)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4569..4592
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4608..4634
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    4658..5209
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5221..5820
                     /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    5837..6425
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6308..6319)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6500..6733
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        6938..7042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7043..7903
                     /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(8074..8357,1..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 ttggcatggt gggccccgcg ccgcggcggc
     4021 ggctgcggcc gctggcagcg ctggccctgg tcctggcgct ggccccgggg ctgcccacag
     4081 cccgggccgg gcagacaccg cgccctgccg agcgggggcc cccagtgcgg cttttcaccg
     4141 aggaggagct ggcccgctat ggcggggagg aggaagatca gcccatctac ttggcagtga
     4201 agggagtggt gtttgatgtc acctccggaa aggagttcta cggcagagga gccccctaca
     4261 atgccttgac ggggaaggac tccactagag gggtagccaa gatgtccttg gatcctgcag
     4321 acctcaccca tgacactacg ggtctcacgg ccaaggaact ggaggccctg gatgaggtct
     4381 tcaccaaagt gtacaaagcc aaatacccca tcgtcggcta cactgcccgg agaattctca
     4441 atgaggatgg cagccctaac ctggacttca agcctgaaga ccagccccat tttgacatca
     4501 aggatgagtt ctgcccaact ttcttgtaca aagtggttga tctagagggc ccgcggttcg
     4561 aactcgagga ctacaaggac gacgatgaca agtcggccgc tggaggatac ccctacgacg
     4621 tgcccgacta cgcctagaat tccgcccccc ccccctaacg ttactggccg aagccgcttg
     4681 gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc
     4741 aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc
     4801 cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa
     4861 gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct
     4921 ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca
     4981 caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca
     5041 agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat
     5101 ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa cgtctaggcc
     5161 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5221 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     5281 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5341 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5401 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5461 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5521 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5581 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5641 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     5701 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     5761 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     5821 tgcatcgatg gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     5881 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     5941 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6001 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6061 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6121 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6181 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6241 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6301 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6361 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6421 cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6481 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     6541 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6601 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6661 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6721 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     6781 aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg
     6841 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     6901 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     6961 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7021 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7081 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7141 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7201 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7261 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     7321 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7381 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7441 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7501 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7561 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7621 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7681 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     7741 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     7801 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     7861 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     7921 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     7981 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8041 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     8101 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     8161 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     8221 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     8281 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     8341 cgggttggac tcaagac
//
