LOCUS       Exported                8841 bp ds-DNA     circular SYN 10-JUN-2020
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8841)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 10, 2020 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8841
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    198..1236
                     /label=TCG folder 292641 (sample 7)
     CDS             262..327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             337..357
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             370..390
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             403..420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             433..456
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             493..516
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             529..576
                     /codon_start=1
                     /label=Beacon
                     /translation="ELFNLLGENQPPVVIK"
     misc_feature    574..1540
                     /label=TCAG folder 292641 (sample 15)
     CDS             592..1491
                     /codon_start=1
                     /label=NHEJ1-SS262/263AA
                     /translation="MEELEQGLLMQPWAWLQLAENSLLAKVFITKQGYALLVSDLQQVW
                     HEQVDTSVVSQRAKELNKRLTAPPAAFLCHLDNLLRPLLKDAAHPSEATFSCDCVADAL
                     ILRVRSELSGLPFYWNFHCMLASPSLVSQHLIRPLMGMSLALQCQVRELATLLHMKDLE
                     IQDYQESGATLIRDRLKTEPFEENSFLEQFMIEKLPEACSIGDGKPFVMNLQDLYMAVT
                     TQEVQVGQKHQGAGDPHTSNSASLQGIDSQCVNQPEQLVAAAPTLSAPEKESTGTSGPL
                     QRPQLSKVKRKKPRGLFS"
     misc_feature    1551..1668
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        1717..2227
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2249..2848
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             2976..3209
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3281..3402
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3442..3577
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3598..3616)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3626..3642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3784..4239
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4265..4369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4370..5230
                     /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"
     rep_origin      5401..5989
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6277..6298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6313..6343
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6351..6367
                     /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     6375..6391
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6412..6430
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6458..6684
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6685..6865
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6912..7037
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7530..7763
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7948..7992
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        8538..8841
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtg ctagcaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
      301 atcgattaca aggatgacga tgacaagcct gcaggagaaa atctgtactt ccaggggaga
      361 ggaggcagcg agaacctgta ctttcagggg cctgcaggaa gacatcacca tcaccatcac
      421 agcggcggaa gatggagcca ccctcagttc gagaagggag gaggaagcgg cggaggcagc
      481 ggaggaggaa gctggagcca cccgcagttc gagaaagcga tcgccagaga gctgttcaac
      541 ctgctgggcg agaaccagcc ccccgtggtg atcaaggcga tcgccaccgg tatggaagaa
      601 ctggagcaag gcctgttgat gcagccatgg gcgtggctac agcttgcaga gaactccctc
      661 ttggccaagg tttttatcac caagcagggc tatgccttgt tggtttcaga tcttcaacag
      721 gtgtggcatg aacaggtgga cactagtgtg gtcagccagc gagccaagga gctgaacaag
      781 cggctcactg ctcctcctgc agctttcctc tgtcatttgg ataatctcct tcgcccattg
      841 ttgaaggacg ctgctcaccc tagcgaagct accttctcct gtgattgtgt ggcagatgca
      901 ctgattctac gggtgcgaag tgagctctct ggcctcccct tctattggaa tttccactgc
      961 atgctagcta gtccttccct ggtctcccaa catttgattc gtcctctgat gggcatgagt
     1021 ctggcattac agtgccaagt gagggagcta gcaacgttac ttcatatgaa agacctagag
     1081 atccaagact accaggagag tggggctacg ctgattcgag atcgattgaa gacagaacca
     1141 tttgaagaaa attccttctt ggaacaattt atgatagaga aactgccaga ggcatgcagc
     1201 attggtgatg gaaagccctt tgtcatgaat ctgcaggatc tgtatatggc agtcaccaca
     1261 caagaggtcc aagtgggaca gaagcatcaa ggcgctggag atcctcatac ctcaaacagt
     1321 gcttccctgc aaggaatcga tagccaatgt gtaaaccagc cagaacaact ggtcgccgcc
     1381 gccccaaccc tctcagcacc tgagaaagag tccacgggta cttcaggccc tctgcagaga
     1441 cctcagctgt caaaggtcaa gaggaagaag ccaaggggtc tcttcagtta atctagatgg
     1501 ggtggcttcg aattctcgac ctcgagacaa atggcagtat tcatccacaa ttttaaaaga
     1561 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1621 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     1681 agggacagca gagatccact ttggccgcgg ctcgaggggg ttggggttgc gccttttcca
     1741 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     1801 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     1861 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     1921 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     1981 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     2041 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     2101 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     2161 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     2221 tccccagggg gatccaccgg agcttaccat gaccgagtac aagcccacgg tgcgcctcgc
     2281 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     2341 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     2401 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     2461 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     2521 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     2581 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     2641 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     2701 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     2761 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     2821 gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac gacccgcagc gcccgaccga
     2881 aaggagcgca cgaccccatg catcggtacc tttaagacca atgacttaca aggcagctgt
     2941 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
     3001 aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg
     3061 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3121 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3181 cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta
     3241 tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta
     3301 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3361 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct
     3421 atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3481 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3541 ctattccaga agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct
     3601 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3661 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3721 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3781 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3841 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3901 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3961 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4021 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4081 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4141 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4201 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     4261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4381 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4441 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4501 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4561 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4621 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4681 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4741 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4801 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4861 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4921 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4981 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5041 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5101 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5161 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5221 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5281 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5341 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5401 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5461 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5521 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5581 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5641 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5701 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5761 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5821 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5881 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5941 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6001 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6061 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     6121 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     6181 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     6241 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     6301 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     6361 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     6421 tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg
     6481 tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac
     6541 cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag
     6601 acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt
     6661 aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag
     6721 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6781 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6841 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     6901 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     6961 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     7021 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     7081 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     7141 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     7201 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     7261 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     7321 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     7381 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     7441 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     7501 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     7561 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     7621 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     7681 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     7741 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     7801 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     7861 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     7921 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     7981 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     8041 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     8101 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     8161 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     8221 attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg agcggccgcc
     8281 cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     8341 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
     8401 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     8461 gactatcata tgttgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     8521 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8581 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     8641 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8701 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8761 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     8821 ttagtcatcg ctattaccat g
//
