LOCUS Exported 8933 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION lentiviral backbone. ACCESSION . VERSION . KEYWORDS pLV-EF1a-IRES-Neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8933) AUTHORS Hayer A, Shao L, Chung M, Joubert LM, Yang HW, Tsai FC, Bisaria A, Betzig E, Meyer T TITLE Engulfed cadherin fingers are polarized junctional structures between collectively migrating endothelial cells. JOURNAL Nat Cell Biol. 2016 Dec;18(12):1311-1323. doi: 10.1038/ncb3438. Epub 2016 Nov 14. PUBMED 27842057 REFERENCE 2 (bases 1 to 8933) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8933 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /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 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" promoter 2221..2432 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 2433..3371 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3328..3348 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind complement(3676..3693) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3903..3922 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 4084..4878 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4138..4157) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4748..4767 /label=Neo-F /note="Neomycin resistance gene, forward primer" misc_feature 4902..5490 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4955..4975) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5373..5384) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5697..6330 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(6458..6474) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6458..6474) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6471..6493) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6482..6498 /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)." promoter complement(6506..6536) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6551..6572 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6689..6706) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6860..7448) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6940..6959) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7619..8479) /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 8242..8261 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8480..8584) /gene="bla" /label=AmpR promoter polyA_signal 8632..8766 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8669..8688) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8723..8742 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgatgaggcc ctttcgtctt cactcgaggt gcccgtcagt 2221 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 2281 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 2341 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 2401 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 2461 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt 2521 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg 2581 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc 2641 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca 2701 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg 2761 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc 2821 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg 2881 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta 2941 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat 3001 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc 3061 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat 3121 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga 3181 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt 3241 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc 3301 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc 3361 ttccatttca ggtgtcgtga ggatccgcgg ccgcgccggc tctagatcgc gaacgcgtga 3421 attcctcgag ggcggccgct ctagagtcga cgggccgcgg taacaattgt taactaactt 3481 aagctagcaa cggtttccct ctagcgggat caattccgcc ccccccccct aacgttactg 3541 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat 3601 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc 3661 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 3721 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 3781 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac 3841 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca 3901 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt 3961 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa 4021 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat 4081 accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4261 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc 4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4861 cttgacgagt tcttctgacc cgggcggggc gcgtctggaa caatcaacct ctggattaca 4921 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 4981 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 5041 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 5101 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 5161 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 5221 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 5281 tggtgttgtc ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga 5341 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 5401 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 5461 gtcggatctc cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga 5521 aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct 5581 agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc 5641 aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga 5701 agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca 5761 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac 5821 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa 5881 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag 5941 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct 6001 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg 6061 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg 6121 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct 6181 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag 6241 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac 6301 ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt 6361 atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt 6421 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa 6481 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct 6541 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc 6601 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg 6661 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 6721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 6781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 7021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 7081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 7141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 7201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 7261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 7321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 7381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 7501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 7561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 7621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 7681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 7741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 7801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 7861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 7921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 7981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 8041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 8101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 8161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 8221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 8281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 8341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 8401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 8461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 8521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 8581 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat caacttgttt 8641 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 8701 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 8761 tggatcaact ggataactca agctaaccaa aatcatccca aacttcccac cccataccct 8821 attaccactg ccaattacct gtggtttcat ttactctaaa cctgtgattc ctctgaatta 8881 ttttcatttt aaagaaattg tatttgttaa atatgtacta caaacttagt agt //