LOCUS       sequence-403766-        7944 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human long non-coding RNA AERRIE 
            (linc01013).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7944)
  AUTHORS   Pham TP, Bink DI, Stanicek L, van Bergen A, van Leeuwen E, Tran Y, 
            Matic L, Hedin U, Wittig I, Dimmeler S, Boon RA
  TITLE     Long Non-coding RNA Aerrie Controls DNA Damage Repair via YBX1 to 
            Maintain Endothelial Cell Function.
  JOURNAL   Front Cell Dev Biol. 2021 Jan 11;8:619079. doi: 
            10.3389/fcell.2020.619079. eCollection 2020.
  PUBMED    33505972
REFERENCE   2  (bases 1 to 7944)
  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: "Front Cell 
            Dev Biol."; date: "2021-01-11"; volume: "8:619079. doi"; pages: " 
            10.3389/fcell.2020.619079. eCollection 2020"
COMMENT     Sequence Label: pLenti4/V5-DEST-AERRIE
FEATURES             Location/Qualifiers
     source          1..7944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(20..44)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             97..138
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(175..195)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        180..509
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      360..495
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        521..568
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             587..961
                     /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             1054..1287
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    1359..1480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1396..1415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1450..1469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1520..1655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1675..1694)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1676..1694)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1704..1721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1704..1720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1713..1735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1862..2317
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1949..1968)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2159..2180
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2343..2447
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2448..2516,2517..3308)
                     /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     complement(2666..2685)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3479..4067
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3968..3987
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4221..4238
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4355..4376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4391..4396,4397..4414,4415..4421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4429..4445
                     /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     4434..4456
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4453..4469
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4453..4469
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4488..4508
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4490..4508
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4536..4762
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4763..4943
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4990..5115
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    5608..5841
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6026..6070
                     /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        6652..6855
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6806..6826
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6857..6875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6878..6896
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6937..6961
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 aagcaagata ttatttgaaa cccagctttc ttgtacaaag tggttgatat ccagcacagt
       61 ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc
      121 ctcggtctcg attctacgcg taccggttag taatgagttt ggaattaatt ctgtggaatg
      181 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      241 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
      301 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
      361 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
      421 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
      481 gcttttttgg aggcctaggc ttttgcaaaa agctccccct gttgacaatt aatcatcggc
      541 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagttgac
      601 cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga
      661 ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga
      721 cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg
      781 ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa
      841 cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga
      901 gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg
      961 acacgtgcta cgagatttaa atggtacctt taagaccaat gacttacaag gcagctgtag
     1021 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     1081 gacaagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     1141 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     1201 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     1261 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     1321 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     1381 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1441 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     1501 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1561 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     1621 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     1681 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     1801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     1861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2281 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     2341 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2401 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2461 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2521 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2581 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2641 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2701 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2761 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2821 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2881 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     2941 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3001 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3061 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3121 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3181 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3241 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3301 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3361 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3421 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3481 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3541 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3601 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3661 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3721 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3781 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3841 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3901 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3961 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4021 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4081 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4141 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4201 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4321 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     4381 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     4441 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     4501 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
     4561 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
     4621 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
     4681 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
     4741 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
     4801 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     4861 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     4921 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     4981 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     5041 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     5101 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     5161 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     5221 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     5281 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     5341 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     5401 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     5461 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     5521 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     5581 gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc
     5641 aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc
     5701 tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt
     5761 gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata
     5821 cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac
     5881 tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac
     5941 gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat
     6001 tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg
     6061 ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa aattattcat
     6121 aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa
     6181 tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg
     6241 acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat
     6301 tcgattagtg aacggatctc gacggtatcg ataagcttgg gagttccgct tacataactt
     6361 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6421 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6481 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     6541 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     6601 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     6661 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     6721 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     6781 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     6841 tatataagca gagctctccc tatcagtgat agagatctcc ctatcagtga tagagatcgt
     6901 cgactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa aaaagcaggc
     6961 tcaggaagcc agcattttta ataaatagga agtaaattat atttaatgac taaattctca
     7021 cctaaagttg atactaatat tacaataact ctcttgtgcc ctattttggt gttttaaact
     7081 tctttgctgt acatttaaga aaatcagaat ttcagtagac tcctcgtgca gtctgaggag
     7141 gcagtttcct atctggtttg acttaagggg ctgcagaaga cattcttatc ccacatattt
     7201 cttctgctga tcagtcagca actgaaattc caactaatgg attccattct tccagttaag
     7261 acatcatctc cagaaaactg tgctagttgg catccagcag tgcccaagta ttatttaacc
     7321 cactagaacc tgtgactgca tcagctttat atcctggacc tcagcctgga acaaaggaga
     7381 gctacacttg gatactaaac acaagtggtc tggatagatt atgctgggaa aagaagcctg
     7441 gagcaggcag tacagtggga gaacaagaca gaagtgcttg ccttggtgat ccaaaaaagg
     7501 caggatccgg ccgggggcaa ggtgattatt ctgtctgatg aggagaataa gagccagttt
     7561 gtggtattga cagaagaatg aaagtaaagt tcaaccaatc tacaaaggcc aaaaatattt
     7621 tcacaggccc ccaaaagatg aactccaatc tgctgcccct aaacgcattc tccactgtct
     7681 ggaagatgtg caaaggcaac ggcacagcgt tctccatctc aaatccttca aagtgctgtc
     7741 agaataaaag aatccccacc aaaagcccca gggaaatcta tctgtctgta tggtatggta
     7801 atgactgagg ttattccaga taggattctg ggagaaactc agctattgaa atctaatgaa
     7861 tggaactgag aatttcttgt tataatactg tttgtttgat tgaatatatc cttgcatata
     7921 ggaagacata aaaataaata ataa
//
