LOCUS       Exported                8415 bp ds-DNA     circular SYN 23-APR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8415)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Dec 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8415
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    679..796
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    854..881
                     /label=bGH
     primer_bind     complement(871..893)
                     /label=Primer 4
     polyA_signal    882..1078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1079
                     /label=Barcode_Cloning_Site
     misc_feature    1166
                     /label=Linker
     misc_feature    1167..1186
                     /label=BFP_Perturb_Primer
     primer_bind     complement(1168..1199)
                     /label=Oligo_19
     misc_feature    1187
                     /label=GFP homology
     primer_bind     1196..1214
                     /label=Oligo_41
     primer_bind     1200..1214
                     /label=Oligo_42
     primer_bind     1200..1209
                     /label=Primer 1
     misc_feature    1201..1789
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1307..1318
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1716..1736
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1778..1824)
                     /label=Oligo_40
     primer_bind     complement(1778..1802)
                     /label=Primer 2
     CDS             complement(1824..2543)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2121..2132)
                     /label=Oligo_19
     misc_feature    2581..2627
                     /label=chimeric intro
     intron          2628..3589
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     promoter        complement(3590..3865)
                     /label=chicken beta-actin promoter
     primer_bind     3631..3711
                     /label=Oligo_131
     primer_bind     3745..3754
                     /label=Oligo_131
     enhancer        3868..4232
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    4233..4245
                     /label=CMV_enhancer
     misc_feature    4336..4655
                     /label=U3
     LTR             4656..4836
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4898..5486)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4978..4997)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5657..6517)
                     /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"
     primer_bind     6280..6299
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6518..6622)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6697..6716)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6888..7267
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7269..7467
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             7485..7665
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7712..7837
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(8334..8415,1..152)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
       61 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
      121 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
      181 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
      241 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
      301 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
      361 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
      421 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
      481 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
      541 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
      601 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
      661 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      721 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      781 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
      841 gggcccgctc tagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
      901 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
      961 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     1021 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagct
     1081 cgagtagaca tgtcgagcta gccaatgatc atacggcgcg ccgaaggtac ccttggatcc
     1141 ctacctaggt gggaattcga taattcgctt gtgccccagt ttgctaggga ggtccaggac
     1201 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     1261 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     1321 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     1381 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     1441 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     1501 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     1561 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     1621 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     1681 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     1741 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgattg gatcgtacgt
     1801 cggcataagc tttgtagcgg ccgctacttg tacagctcgt ccatgccgag agtgatcccg
     1861 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     1921 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1981 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     2041 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     2101 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     2161 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     2221 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     2281 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     2341 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     2401 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     2461 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     2521 agctcctcgc ccttgctcac catggtggcg accggtagcg ctagttacga ctagcaggag
     2581 ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac
     2641 acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccctgcgg
     2701 cagccccgag gctggagatg gagaagggga cggcggcgcg gcgacgcacg aaggccctcc
     2761 ccgcccattt ccttcctgcc ggcgccgcac cgcttcgccc gcgcccgcta gagggggtgc
     2821 ggcggcgcct cccagatttc ggctccgcca gatttgggac aaaggaagtc cctgcgccct
     2881 ctcgcacgat taccataaaa ggcaatggct gcggctcgcc gcgcctcgac agccgccggc
     2941 gctccggggc cgccgcgccc ctcccccgag ccctccccgg cccgaggcgg ccccgccccg
     3001 cccggcaccc ccacctgccg ccaccccccg cccggcacgg cgagccccgc gccacgcccc
     3061 gcacggagcc ccgcacccga agccgggccg tgctcagcaa ctcggggagg ggggtgcagg
     3121 ggggggttac agcccgaccg ccgcgcccac accccctgct caccccccca cgcacacacc
     3181 ccgcacgcag cctttgttcc cctcgcagcc cccccgcacc gcggggcacc gcccccggcc
     3241 gcgctcccct cgcgcacacg cggagcgcac aaagccccgc gccgcgcccg cagcgctcac
     3301 agccgccggg cagcgcgggc cgcacgcggc gctccccacg cacacacaca cgcacgcacc
     3361 ccccgagccg ctcccccccg cacaaagggc cctcccggag ccctttaagg ctttcacgca
     3421 gccacagaaa agaaacgagc cgtcattaaa ccaagcgcta attacagccc ggaggagaag
     3481 ggccgtcccg cccgctcacc tgtgggagta acgcggtcag tcagagccgg ggcgggcggc
     3541 gcgaggcggc gcggagcggg gcacggggcg aaggcaacgc agcgactccc gcccgccgcg
     3601 cgcttcgctt tttatagggc cgccgccgcc gccgcctcgc cataaaagga aactttcgga
     3661 gcgcgccgct ctgattggct gccgccgcac ctctccgcct cgccccgccc cgcccctcgc
     3721 cccgccccgc cccgcctggc gcgcgccccc cccccccccg cccccatcgc tgcacaaaat
     3781 aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat ggggagagtg
     3841 aagcagaacg tggggctcac ctcgacccat ggtaatagcg atgactaata cgtagatgta
     3901 ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg cgggccattt
     3961 accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat gtactgccaa
     4021 gtgggcagtt taccgtaaat agtccaccca ttgacgtcaa tggaaagtcc ctattggcgt
     4081 tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc
     4141 aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc tatgaactaa
     4201 tgaccccgta attgattact attaataact agtcaataat caatgtcgag ctgatctacg
     4261 atgtagtacc tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa
     4321 aagaaaaggg gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc
     4381 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
     4441 ggatcagata tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga
     4501 aggtagaaga agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg
     4561 ggatggatga cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc
     4621 atcacatggc ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct
     4681 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4741 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4801 tcagaccctt ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa ggccagcaaa
     4861 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4921 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4981 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5041 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5101 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5161 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5221 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5281 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5341 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5401 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5461 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5521 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5581 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5641 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5701 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5761 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5821 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5881 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5941 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6001 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6061 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6121 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6181 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6241 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6301 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6361 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6421 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6481 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6541 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6601 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6661 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     6721 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     6781 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     6841 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     6901 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6961 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     7021 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     7081 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     7141 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     7201 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     7261 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     7321 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     7381 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     7441 tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct ctctggttag
     7501 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     7561 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     7621 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
     7681 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
     7741 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     7801 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     7861 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     7921 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     7981 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     8041 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     8101 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     8161 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     8221 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     8281 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     8341 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     8401 acggtacagg ccaga
//
