LOCUS       sequence-383226-        7368 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Lentiviral vector expressing long non-coding RNA LASSIE (linc00520).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7368)
  AUTHORS   Stanicek L, Lozano-Vidal N, Bink DI, Hooglugt A, Yao W, Wittig I, 
            van Rijssel J, van Buul JD, van Bergen A, Klems A, Ramms AS, Le 
            Noble F, Hofmann P, Szulcek R, Wang S, Offermanns S, Ercanoglu MS, 
            Kwon HB, Stainier D, Huveneers S, Kurian L, Dimmeler S, Boon RA
  TITLE     Long non-coding RNA LASSIE regulates shear stress sensing and 
            endothelial barrier function.
  JOURNAL   Commun Biol. 2020 May 26;3(1):265. doi: 10.1038/s42003-020-0987-0.
  PUBMED    32457386
REFERENCE   2  (bases 1 to 7368)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2020-05-26"; volume: "3(1):265. doi"; pages: " 
            10.1038/s42003-020-0987-0"
COMMENT     Sequence Label: pLenti4/V5-DEST-LASSIE
FEATURES             Location/Qualifiers
     source          1..7368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(115..134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      928..1516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1417..1436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1670..1687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1804..1825
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1840..1845,1846..1863,1864..1870)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1878..1894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1883..1905
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1902..1918
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1902..1918
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1937..1957
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1939..1957
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1985..2211
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2212..2392
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2439..2564
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3057..3290
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3475..3519
                     /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"
     promoter        4101..4304
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4255..4275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4306..4324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4327..4345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4386..4410
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4837..4861)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4914..4955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(4992..5012)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4997..5326
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5177..5312
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        5338..5385
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5404..5778
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             5871..6104
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6176..6297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6213..6232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6267..6286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6337..6472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6492..6511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6493..6511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6521..6538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6521..6537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6530..6552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6679..7134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6766..6785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6976..6997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7160..7264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7265..7333,7334..7368,1..757)
                     /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"
ORIGIN
        1 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
       61 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      121 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      181 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      241 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      301 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      361 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      421 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      481 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      541 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      601 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      661 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      721 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
      781 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
      841 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
      901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
      961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1561 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1621 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1681 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     1741 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     1801 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     1861 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     1921 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     1981 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
     2041 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
     2101 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
     2161 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
     2221 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     2281 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     2341 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
     2401 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
     2461 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
     2521 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
     2581 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
     2641 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
     2701 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
     2761 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
     2821 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
     2881 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
     2941 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
     3001 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
     3061 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
     3121 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
     3181 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
     3241 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
     3301 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
     3361 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
     3421 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
     3481 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
     3541 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
     3601 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
     3661 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
     3721 ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg acggtatcga
     3781 taagcttggg agttccgctt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3841 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3901 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3961 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4021 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4081 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     4141 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4201 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4261 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
     4321 gagatctccc tatcagtgat agagatcgtc gactagtcca gtgtggtgga attctgcaga
     4381 tatcaacaag tttgtacaaa aaagcaggct ccgcggccgc ccccttcacc gggtcttgcc
     4441 ttctgccatg taagatatga ctgtgctcct ccttcacctt ctgccatgat tgtgaggcct
     4501 ccacagccat gtggaactgg aagctgtttt tccctctttg gggtctcaga gagattggtg
     4561 gcaaggactc ctgggaagag aaaagctgaa ggacactctg gcaagaggaa accccgagcc
     4621 aaacgtatgg ccacctctga ccactcgacc agcccagcat ggtgtacatt tctgggtagc
     4681 ttatcacgcc ttccaagcat tgcctttatc ttccttctcc ccatcaagcc tgtattgttt
     4741 tccttttgaa tcataagcac ttttgttaaa tacctcaaat ctttcgagaa cagcaaggag
     4801 caaaataaac ttaacttctc aagggtgggc gcgccgaccc agctttcttg tacaaagtgg
     4861 ttgatatcca gcacagtggc ggccgctcga gtctagaggg cccgcggttc gaaggtaagc
     4921 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttgga
     4981 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     5041 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     5101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     5161 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     5221 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     5281 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tccccctgtt
     5341 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     5401 accatggcca agttgaccag tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg
     5461 gtcgagttct ggaccgaccg gctcgggttc tcccgggact tcgtggagga cgacttcgcc
     5521 ggtgtggtcc gggacgacgt gaccctgttc atcagcgcgg tccaggacca ggtggtgccg
     5581 gacaacaccc tggcctgggt gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg
     5641 gaggtcgtgt ccacgaactt ccgggacgcc tccgggccgg ccatgaccga gatcggcgag
     5701 cagccgtggg ggcgggagtt cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg
     5761 gccgaggagc aggactgaca cgtgctacga gatttaaatg gtacctttaa gaccaatgac
     5821 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     5881 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     5941 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6001 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6061 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6121 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6301 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     6481 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     6541 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6601 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6661 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6721 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6781 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6841 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6901 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6961 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     7021 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     7081 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     7141 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7201 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7261 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7321 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctga
//
