LOCUS       Exported                9204 bp ds-DNA     circular SYN 27-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9204)
  AUTHORS   David Gordon
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Mar 25, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9204
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /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    2028..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2338..3519
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2568..3510
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon"
     misc_feature    3525..3540
                     /label=5' Homology with pLVX-EF1a-IRES-Puro
     misc_feature    3535
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3541..3549
                     /label=Kozak Consensus Sequence
     CDS             3550..3840
                     /codon_start=1
                     /label=orf9b
                     /translation="MDPKISEMHPALRLVDPQIQLAVTRMENAVGRDQNNVGPKVYPII
                     LRLGSPLSLNMARKTLNSLEDKAFQLTPIAVQMTKLATTEELPDEFVVVTVK"
     CDS             3859..3882
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3919..3942
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     misc_feature    3946..3961
                     /label=3' Homology with pLVX-EF1a-IRES-Puro
     misc_feature    3947..3951
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    3954..4527
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4560..5159
                     /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    5173..5761
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5968..6601
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7131..7719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7890..8750)
                     /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(8751..8855)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgatgagtaa ttcatacaaa aggactcgcc cctgccttgg
     2221 ggaatcccag ggaccgtcgt taaactccca ctaacgtaga acccagagat cgctgcgttc
     2281 ccgccccctc acccgcccgc tctcgtcatc actgaggtgg agaagagcat gcgtgaggct
     2341 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     2401 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     2461 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     2521 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
     2581 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
     2641 ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
     2701 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
     2761 cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
     2821 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
     2881 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
     2941 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
     3001 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
     3061 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     3121 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     3181 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     3241 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     3301 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     3361 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     3421 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     3481 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatctatttc cggtgaattc
     3541 gccgccacca tggaccccaa aatttctgaa atgcacccgg cgcttcgctt ggtggacccc
     3601 caaatccaac tcgctgtcac gcggatggaa aacgcagtag gacgcgacca aaataatgtc
     3661 ggcccaaaag tgtatccgat tatcctccgc cttggcagtc ccctgagcct gaatatggcc
     3721 agaaagactc ttaacagcct ggaggacaaa gcgtttcagc tgacccccat tgcggttcag
     3781 atgactaagc tggcgacaac tgaagagttg cccgacgaat ttgtggtagt aaccgtaaag
     3841 ctcgaaggcg gcgggggatg gagccatcca caattcgaga aaggcggtgg ttcaggagga
     3901 ggtagcgggg gtggatcatg gtcacatccg cagtttgaaa agtaaggatc ccgcccctct
     3961 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     4021 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     4081 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     4141 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     4201 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     4261 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     4321 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     4381 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     4441 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     4501 gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga cgaccttcca
     4561 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc cgggccgtac
     4621 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gacccggacc
     4681 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4741 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4801 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4861 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
     4921 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4981 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     5041 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     5101 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctaga
     5161 cgcgtctgga acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     5221 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     5281 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     5341 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     5401 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     5461 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     5521 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     5581 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     5641 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     5701 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     5761 cctggaatta attctgcagt cgagacctag aaaaacatgg agcaatcaca agtagcaata
     5821 cagcagctac caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt
     5881 ttccagtcac acctcaggta cctttaagac caatgactta caaggcagct gtagatctta
     5941 gccacttttt aaaagaaaag aggggactgg aagggctaat tcactcccaa cgaagacaag
     6001 atatccttga tctgtggatc taccacacac aaggctactt ccctgattag cagaactaca
     6061 caccagggcc aggggtcaga tatccactga cctttggatg gtgctacaag ctagtaccag
     6121 ttgagccaga taaggtagaa gaggccaata aaggagagaa caccagcttg ttacaccctg
     6181 tgagcctgca tgggatggat gacccggaga gagaagtgtt agagtggagg tttgacagcc
     6241 gcctagcatt tcatcacgtg gcccgagagc tgcatccgga gtacttcaag aactgctgat
     6301 atcgagcttg ctacaaggga ctttccgctg gggactttcc agggaggcgt ggcctgggcg
     6361 ggactgggga gtggcgagcc ctcagatcct gcatataagc agctgctttt tgcctgtact
     6421 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     6481 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     6541 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     6601 agtagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa atgaatatca
     6661 gagagtgaga ggccttgaca ttgctagcgt ttaccgtcga cctctagcta gagcttggcg
     6721 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6781 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6841 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6901 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6961 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7021 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7081 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     7141 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     7201 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7261 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7321 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7381 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7441 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7501 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7561 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7621 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7681 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7741 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7801 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7861 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7921 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7981 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8041 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     8101 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     8161 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     8221 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8281 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8341 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8401 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8461 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8521 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8581 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8641 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8701 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8761 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8821 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8881 gacgtcgacg gatcgggaga tcaacttgtt tattgcagct tataatggtt acaaataaag
     8941 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     9001 gtccaaactc atcaatgtat cttatcatgt ctggatcaac tggataactc aagctaacca
     9061 aaatcatccc aaacttccca ccccataccc tattaccact gccaattacc tgtggtttca
     9121 tttactctaa acctgtgatt cctctgaatt attttcattt taaagaaatt gtatttgtta
     9181 aatatgtact acaaacttag tagt
//
