LOCUS       sequence-372800-        7706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lenticiral expression of EGFP-HRasC20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7706)
  AUTHORS   Katsuno-Kambe H, Teo JL, Ju RJ, Hudson J, Stehbens SJ, Yap AS
  TITLE     Collagen polarization promotes epithelial elongation by stimulating 
            locoregional cell proliferation.
  JOURNAL   Elife. 2021 Oct 18;10. pii: 67915. doi: 10.7554/eLife.67915.
  PUBMED    34661524
REFERENCE   2  (bases 1 to 7706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-10-18"; volume: "10. pii: 67915. doi"; pages: " 
            10.7554/eLife.67915"
COMMENT     Sequence Label: pLL5.0-EGFP-HRasC20
FEATURES             Location/Qualifiers
     source          1..7706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(72..91)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        263..642
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        644..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     797..817
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             860..1040
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1087..1212
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1709..1942
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2127..2171
                     /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    2469..2586
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2641..2953
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2872..2892
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2970..2986
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3028..3061)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3656..3665
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3662..3664,3665..3667,3668..4378)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3662..3664,3665..3667,3668..4378)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3707..3728)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3968..3987)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4315..4336
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(4533..4566)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4622..5208
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4675..4695)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5093..5104)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5211..5227)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5737..5917
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5979..6567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6059..6078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6738..7529,7530..7598))
                     /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     7361..7380
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7599..7703)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
       61 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
      121 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
      181 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      241 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      301 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      361 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      421 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      481 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      541 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      601 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      661 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      721 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      781 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      841 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
      901 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
      961 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     1021 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     1081 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     1141 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     1201 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     1261 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     1321 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     1381 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     1441 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     1501 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc
     1561 tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa ttatataaat
     1621 ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag agaagagtgg
     1681 tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc ttgggagcag
     1741 caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga caattattgt
     1801 ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa cagcatctgt
     1861 tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct gtggaaagat
     1921 acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc atttgcacca
     1981 ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt tggaatcaca
     2041 cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata cactccttaa
     2101 ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa ttagataaat
     2161 gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata aaattattca
     2221 taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt tctatagtga
     2281 atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca accccgaggg
     2341 gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga gacagatcca
     2401 ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat ggcagtattc
     2461 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     2521 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     2581 aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc gcgctctaga
     2641 gatccgacgc gccatctcta ggcccgcgcc ggccccctcg cacagacttg tgggagaagc
     2701 tcggctactc ccctgccccg gttaatttgc atataatatt tcctagtaac tatagaggct
     2761 taatgtgcga taaaagacag ataatctgtt ctttttaata ctagctacat tttacatgat
     2821 aggcttggat ttctataaga gatacaaata ctaaattatt attttaaaaa acagcacaaa
     2881 aggaaactca ccctaactgt aaagtaattg tgtgttttga gactataaat atcccttgga
     2941 gaaaagcctt gttaacgcgc ggtgaccctc gaggtcgacg gtatcgataa gctcgcttca
     3001 cgagattcca gcaggtcgag ggacctaata acttcgtata gcatacatta tacgaagtta
     3061 tattaagggt tccaagctta agcggccgct gaaagacccc acctgtaggt ttggcaagct
     3121 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     3181 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     3241 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     3301 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     3361 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     3421 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     3481 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     3541 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     3601 catcgaattg cagagctggt ttagtgaacc gtcagatccg ctagcgctac cggtcgccac
     3661 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     3721 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     3781 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     3841 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     3901 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     3961 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     4021 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     4081 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     4141 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     4201 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     4261 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     4321 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagtc
     4381 cggactcaga tctaagctga accctcctga tgagagtggc cccggctgca tgagctgcaa
     4441 gtgtgtgctc tcctgaggat ccaccggatc tagataactg atcataatca gccataccac
     4501 atttgtagag gggcaattcg tcgagggacc taataacttc gtatagcata cattatacga
     4561 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     4621 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     4681 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     4741 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     4801 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     4861 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     4921 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     4981 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     5041 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     5101 caatccagcg gaccttcctt ccaggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5161 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     5221 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     5281 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     5341 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     5401 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     5461 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     5521 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     5581 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     5641 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     5701 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     5761 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     5821 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     5881 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     5941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6661 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6721 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6781 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6841 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6901 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6961 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7021 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7081 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7141 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7201 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7261 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7321 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7381 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7441 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7501 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7561 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7621 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7681 aataaacaaa taggggttcc gcgcac
//
