LOCUS       Exported                7652 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral expression of K-Ras C14.
ACCESSION   .
VERSION     .
KEYWORDS    pLL5.0 mCherry_K-RasC14
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7652)
  AUTHORS   Wu SK, Gomez GA, Michael M, Verma S, Cox HL, Lefevre JG, Parton RG, 
            Hamilton NA, Neufeld Z, Yap AS
  TITLE     Cortical F-actin stabilization generates apical-lateral patterns of 
            junctional contractility that integrate cells into epithelia.
  JOURNAL   Nat Cell Biol. 2014 Feb;16(2):167-78. doi: 10.1038/ncb2900. Epub 
            2014 Jan 12.
  PUBMED    24413434
REFERENCE   2  (bases 1 to 7652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7652
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        33..412
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        414..612
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     567..587
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             630..810
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    857..982
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1479..1712
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1897..1941
                     /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    2239..2356
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2642..2662
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2740..2756
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2798..2831
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      3419..3428
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(3570..3588)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3838..3857
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    4249..4282
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    4338..4924
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4391..4411)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4809..4820)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4927..4943)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5453..5633
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5695..6283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5775..5794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6454..7314)
                     /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     7077..7096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7315..7419)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7494..7513)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
       61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      421 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      541 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      601 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      661 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      721 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      781 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      841 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      901 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      961 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     1021 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1081 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1141 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1201 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1261 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1321 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1381 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1441 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1501 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1561 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1621 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1681 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1741 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1801 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1861 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1921 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1981 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     2041 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     2101 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     2161 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
     2221 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
     2281 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
     2341 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
     2401 gcgctctaga gatccgacgc gccatctcta ggcccgcgcc ggccccctcg cacagacttg
     2461 tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac
     2521 tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat
     2581 tttacatgat aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa
     2641 acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat
     2701 atcccttgga gaaaagcctt gttaacgcgc ggtgaccctc gaggtcgacg gtatcgataa
     2761 gctcgcttca cgagattcca gcaggtcgag ggacctaata acttcgtata gcatacatta
     2821 tacgaagtta tattaagggt tccaagctta agcggccgct gaaagacccc acctgtaggt
     2881 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     2941 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     3001 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     3061 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     3121 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     3181 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     3241 ccgatagact gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc
     3301 cgaatcgtgg actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc
     3361 gggggtcttt catcgaattc tgcagtcgac ggtaccgcgg gcccgggatc caccggtcgc
     3421 caccatggtg agcatggacg aggaggataa catggccatc atcaaggagt tcatgcgctt
     3481 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     3541 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     3601 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     3661 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     3721 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     3781 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     3841 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     3901 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     3961 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     4021 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     4081 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca aaaagaagaa
     4141 aaagaagtca aagacaaagt gtgtaattat gtaagtacaa gtaaagcggc cgcgactcta
     4201 gatcataatc agccatacca ccctgcaggc aattcgtcga gggacctaat aacttcgtat
     4261 agcatacatt atacgaagtt atacatgttt aagggttccg gttccactag gtacaattcg
     4321 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     4381 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     4441 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     4501 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     4561 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     4621 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     4681 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     4741 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct
     4801 acgtcccttc ggccctcaat ccagcggacc ttccttcccg gcctgctgcc ggctctgcgg
     4861 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4921 ccgcatcgat accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag
     4981 caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt
     5041 gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga
     5101 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
     5161 acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa
     5221 ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt
     5281 accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca
     5341 ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga
     5401 cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct
     5461 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     5521 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     5581 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg
     5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     7021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     7081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     7141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7441 acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc
     7501 tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct
     7561 gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg
     7621 aagaatctgc ttagggttag gcgttttgcg ct
//
