LOCUS       Exported                9435 bp ds-DNA     circular SYN 02-DEC-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pNL(CMV)Gluc/CMV/WPRE-Delta-U3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 2, 2019 from SnapGene Viewer 4.3.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9435
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(941..1801)
                     /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(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3324..3504
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      4131..4266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4544..4777
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     enhancer        5276..5579
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5580..5783
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5909..5925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5948..5966
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     protein_bind    5991..6015
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     propeptide      6056..6127
     misc_feature    6128..6634
                     /label=GLuc
     misc_feature    6635..6802
                     /label=CD8_TM
     protein_bind    complement(6829..6853)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(6885..6903)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6914..6930)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6962..7550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(7433..7444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7625..7858
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     1981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     2041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     2101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     2161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2281 cggtgcgggc ctcttcgcta ttacgccagg ggaggcagag attgcagtaa gctgagatcg
     2341 cagcactgca ctccagcctg ggcgacagag taagactctg tctcaaaaat aaaataaata
     2401 aatcaatcag atattccaat cttttccttt atttatttat ttattttcta ttttggaaac
     2461 acagtccttc cttattccag aattacacat atattctatt tttctttata tgctccagtt
     2521 ttttttagac cttcacctga aatgtgtgta tcgagaccta gaaaaacatg gagcaatcac
     2581 aagtagcaat acagcagcta acaatgctgc ttgtgcctgg ctagaagcac aagaggagga
     2641 agaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt acaaggcagc
     2701 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     2761 aagaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     2821 gcagaactac acaccagggc caggggtcag atatccactg acctttggat ggtgctacaa
     2881 gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga acaccagctt
     2941 gttacaccct gtgagcctgc atggaatgga tgaccctgag agagaagtgt tagagtggag
     3001 gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg agtatctaga
     3061 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     3121 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     3181 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3241 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tactctagat gctacatata
     3301 agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3361 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgctca
     3421 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3481 agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag cgaaagtaaa
     3541 gccagaggag atctctcgac gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga
     3601 ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga aggagagaga
     3661 tgggtgcgag agcgtcggta ttaagcgggg gagaattaga taaatgggaa aaaattcggt
     3721 taaggccagg gggaaagaaa caatataaac taaaacatat agtatgggca agcagggagc
     3781 tagaacgatt cgcagttaat cctggccttt tagagacatc agaaggctgt agacaaatac
     3841 tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata
     3901 caatagcagt cctctattgt gtgcatcaaa ggatagatgt aaaagacacc aaggaagcct
     3961 tagataagat agaggaagag caaaacaaaa gtaagaaaaa ggcacagcaa gcagcagctg
     4021 acacaggaaa caacagccag gtcagccaaa attaccctat agtgcagaac ctccaggggc
     4081 aaatggtaca tcaggccata tcacctagaa ctttaaatat gactccgccc atcccgcccc
     4141 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4201 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4261 gaggcctagg cttttgcaaa tgcataaaag aaaagggggg attggggggt acagtgcagg
     4321 ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac aaaaacaaat
     4381 tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccac aattggagaa
     4441 gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca cccaccaagg
     4501 caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct ttgttccttg
     4561 ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg acggtacagg
     4621 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
     4681 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
     4741 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
     4801 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
     4861 agatttggaa taacatgacc tggatggagt gggacagaga aattaacaat tacacaagct
     4921 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
     4981 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
     5041 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
     5101 tactttctgt agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
     5161 tcccaatccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
     5221 acagagacag atccattcga ttagtgaacg gatctagtta actagatcta gagtccgtta
     5281 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     5341 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     5401 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     5461 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     5521 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     5581 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     5641 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     5701 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5761 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     5821 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg actctagcct
     5881 aggccgcggg acggataaca atttcacaca ggaaacagct atgaccatta ggcctttgca
     5941 aaaagctatt taggtgacac tatagaaggt acgcctgcag gtaccggatc acaagtttgt
     6001 acaaaaaagc aggctggcgc cggaaccaat tcagtcgact ggatccggta ccatcatggc
     6061 cctgcccgtg acagctctgc tgctgcctct ggccctgctg ctgcatgccg ccagacccgg
     6121 gatccaaatg aagcccaccg agaacaacga agacttcaac atcgtggccg tggccagcaa
     6181 cttcgcgacc acggatctcg atgctgaccg cgggaagttg cccggcaaga agctgccgct
     6241 ggaggtgctc aaagagatgg aagccaatgc ccggaaagct ggctgcacca ggggctgtct
     6301 gatctgcctg tcccacatca agtgcacgcc caagatgaag aagttcatcc caggacgctg
     6361 ccacacctac gaaggcgaca aagagtccgc acagggcggc ataggcgagg cgatcgtcga
     6421 catccccgag atccccggct tcaaggacct ggaacccatg gaacagttta tcgcccaggt
     6481 ggacctgtgc gtggactgca ccaccggctg tctgaagggc ctggccaacg tgcagtgcag
     6541 cgacctgctg aagaagtggc tgccccagag atgcgccacc ttcgccagca agatccaggg
     6601 ccaggtggac aagatcaagg gcgctggcgg cgacgcttgt agacctgccg ctggcggagc
     6661 cgtgcacacc agaggcctgg acttcgcctg cgacatctac atctgggccc ctctggccgg
     6721 aacatgtggc gtgctgctgc tgagcctggt catcaccctg tactgcaacc accggaaccg
     6781 gcggagagtg tgcaagtgcc cctgagcggc cgcactcgag atatctagac ccagctttct
     6841 tgtacaaagt ggtgatcgcg tgcatgcgac gtcatagctc tctccctata gtgagtcgta
     6901 ttataagcta ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac tgctagtcga
     6961 gaatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     7021 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     7081 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     7141 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     7201 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     7261 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     7321 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     7381 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     7441 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     7501 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctgggtacct
     7561 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     7621 ggactggaag ggctaattca ctcccaaaga agacaagatc tgctttttgc ctgtactggg
     7681 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     7741 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     7801 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcacc
     7861 caggaggtag aggttgcagt gagccaagat cgcgccactg cattccagcc tgggcaagaa
     7921 aacaagactg tctaaaataa taataataag ttaagggtat taaatatatt tatacatgga
     7981 ggtcataaaa atatatatat ttgggctggg cgcagtggct cacacctgcg cccggccctt
     8041 tgggaggccg aggcaggtgg atcacctgag tttgggagtt ccagaccagc ctgaccaaca
     8101 tggagaaacc ccttctctgt gtatttttag tagattttat tttatgtgta ttttattcac
     8161 aggtatttct ggaaaactga aactgttttt cctctactct gataccacaa gaatcatcag
     8221 cacagaggaa gacttctgtg atcaaatgtg gtgggagagg gaggttttca ccagcacatg
     8281 agcagtcagt tctgccgcag actcggcggg tgtccttcgg ttcagttcca acaccgcctg
     8341 cctggagaga ggtcagacca cagggtgagg gctcagtccc caagacataa acacccaaga
     8401 cataaacacc caacaggtcc accccgcctg ctgcccaggc agagccgatt caccaagacg
     8461 ggaattagga tagagaaaga gtaagtcaca cagagccggc tgtgcgggag aacggagttc
     8521 tattatgact caaatcagtc tccccaagca ttcggggatc agagttttta aggataactt
     8581 agtgtgtagg gggccagtga gttggagatg aaagcgtagg gagtcgaagg tgtccttttg
     8641 cgccgagtca gttcctgggt gggggccaca agatcggatg agccagttta tcaatccggg
     8701 ggtgccagct gatccatgga gtgcagggtc tgcaaaatat ctcaagcact gattgatctt
     8761 aggttttaca atagtgatgt taccccagga acaatttggg gaaggtcaga atcttgtagc
     8821 ctgtagctgc atgactccta aaccataatt tcttttttgt tttttttttt ttatttttga
     8881 gacagggtct cactctgtca cctaggctgg agtgcagtgg tgcaatcaca gctcactgca
     8941 gcctcaacgt cgtaagctca agcgatcctc ccacctcagc ctgcctggta gctgagacta
     9001 caagcgacgc cccagttaat ttttgtattt ttggtagagg cagcgttttg ccgtgtggcc
     9061 ctggctggtc tcgaactcct gggctcaagt gatccagcct cagcctccca aagtgctggg
     9121 acaaccgggg ccagtcactg cacctggccc taaaccataa tttctaatct tttggctaat
     9181 ttgttagtcc tacaaaggca gtctagtccc caggcaaaaa gggggtttgt ttcgggaaag
     9241 ggctgttact gtctttgttt caaactataa actaagttcc tcctaaactt agttcggcct
     9301 acacccagga atgaacaagg agagcttgga ggttagaagc acgatggaat tggttaggtc
     9361 agatctcttt cactgtctga gttataattt tgcaatggtg gttcaaagac tgcccgcttc
     9421 tgacaccagt cgctg
//
