LOCUS       Exported                7563 bp ds-DNA     circular SYN 17-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSico
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L,
            Jaenisch R, Jacks T
  TITLE     Cre-lox-regulated conditional RNA interference from transgenes.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jul 13. 101(28):10380-5.
  PUBMED    15240889
REFERENCE   2  (bases 1 to 7563)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7563
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             342..386
                     /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"
     misc_feature    684..801
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     1091..1111
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    1129..1162
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     enhancer        1262..1565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1566..1769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1808..1817
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1814..2530
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1859..1880)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2120..2139)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2467..2488
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2541..2574
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
     primer_bind     2601..2617
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2692..3280
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2745..2765)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3163..3174)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3283..3299)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3809..3989
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4051..4639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4131..4150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4810..5670)
                     /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     5433..5452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5671..5775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5850..5869)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6041..6420
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6422..6620
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6638..6818
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6865..6990
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7487..157
                     /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 ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg gctattgagg
       61 cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag gcaagaatcc
      121 tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt tgctctggaa
      181 aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa tctctggaac
      241 agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat tacacaagct
      301 taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa caagaattat
      361 tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat tggctgtggt
      421 atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata gtttttgctg
      481 tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt cagacccacc
      541 tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt ggagagagag
      601 acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca attctgcaga
      661 caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg gtacagtgca
      721 ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa
      781 attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc agtttggtta
      841 gtaccgggcc cgctctagag atccgacgcg ccatctctag gcccgcgccg gccccctcgc
      901 acagacttgt gggagaagct cggctactcc cctgccccgg ttaatttgca tataatattt
      961 cctagtaact atagaggctt aatgtgcgat aaaagacaga taatctgttc tttttaatac
     1021 tagctacatt ttacatgata ggcttggatt tctataagag atacaaatac taaattatta
     1081 ttttaaaaaa cagcacaaaa ggaaactcac cctaactgta aagtaattat aacttcgtat
     1141 agtataaatt atacgaagtt ataagccttg gttttttgaa ttccgtatta ccgccatgca
     1201 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     1261 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     1321 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     1381 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     1441 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1501 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1561 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1621 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1681 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1741 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     1801 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     1861 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     1921 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     1981 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2041 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2101 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     2161 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     2221 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     2281 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     2341 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     2401 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     2461 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     2521 gctgtacaag tagcggccgc ataacttcgt atagtataaa ttatacgaag ttataagcct
     2581 tgttaacgcg cggtgaccct cgaggtcgac ggtatcgata agctcgcttc acgagatcat
     2641 gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct
     2701 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     2761 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     2821 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     2881 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     2941 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3001 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3061 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     3121 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3181 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3241 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     3301 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     3361 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     3421 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     3481 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     3541 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     3601 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     3661 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     3721 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     3781 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     3841 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     3901 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     3961 cttttagtca gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag
     4021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     5821 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5881 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     5941 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     6001 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
     6061 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     6121 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6181 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6241 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6301 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6361 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6421 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     6481 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6541 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     6601 tgggaggtct atataagcag cgcgttttgc ctgtactggg tctctctggt tagaccagat
     6661 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6721 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6781 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg
     6841 aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc
     6901 acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc
     6961 tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga
     7021 tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta
     7081 tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa
     7141 ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt
     7201 agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa
     7261 gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca
     7321 cagcaagcgg ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     7381 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     7441 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     7501 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     7561 agg
//
