LOCUS Exported 8909 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses HLA-A. ACCESSION . VERSION . KEYWORDS p7128 pHAGE-P-CMVt-N-HA-GAW-HLA-A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8909) AUTHORS Galligan JT, Martinez-Noel G, Arndt V, Hayes S, Chittenden TW, Harper JW, Howley PM TITLE Proteomic analysis and identification of cellular interactors of the giant ubiquitin ligase HERC2. JOURNAL J Proteome Res. 2015 Feb 6;14(2):953-66. doi: 10.1021/pr501005v. Epub 2014 Dec 15. PUBMED 25476789 REFERENCE 2 (bases 1 to 8909) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8909 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 30..618 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 519..538 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 772..789 /label=L4440 /note="L4440 vector, forward primer" primer_bind 881..900 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 1029..1050 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 1065..1095 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1103..1119 /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 1108..1130 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 1127..1143 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1127..1143 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 1193..1212 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 1404..2037 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 2084..2209 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 2706..2939 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 3124..3168 /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 3461..3578 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 3630..3933 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3934..4137 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 4088..4108 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 4139..4157 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4160..4178 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 4188..4212 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 4299..4308 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 4311..4337 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind 4353..4377 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" protein_bind complement(5473..5497) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter 5516..6015 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5551..5574) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5948..5967 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 6036..6635 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(6036..6055) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6532..6552 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 6702..7290 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6755..6775) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7173..7184) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7365..7598 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7717..7735) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 7803..7907 /gene="bla" /label=AmpR promoter CDS 7908..8768 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8126..8145) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 781 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 841 attcattaat gcagcaagct catggctgac taattttttt tatttatgca gaggccgagg 901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 961 tttgcaaaaa gctccccgtg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 1021 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt 1081 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat 1141 gacatgatta cgaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 1201 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa 1261 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat 1321 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa 1381 atatgtacta caaacttagt agttggaagg gctaattcac tcccaaagaa gacaagatat 1441 ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga actacacacc 1501 agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag taccagttga 1561 gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac accctgtgag 1621 cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg acagccgcct 1681 agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact gctgatatcg 1741 agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc tgggcgggac 1801 tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc tgtactgggt 1861 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc 1921 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg 1981 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg 2041 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac 2101 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa 2161 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg 2221 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata 2281 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg 2341 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca 2401 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca 2461 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa 2521 caaaagtaag accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga 2581 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 2641 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 2701 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 2761 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 2821 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 2881 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 2941 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 3001 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 3061 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 3121 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 3181 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 3241 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 3301 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 3361 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga 3421 cggtatcgcc gaattcacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga 3481 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 3541 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca 3601 gagatccagt ttggactagt ggagttccgc gttacataac ttacggtaaa tggcccgcct 3661 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 3721 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 3781 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 3841 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 3901 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 3961 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 4021 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 4081 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc 4141 cctatcagtg atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg 4201 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg 4261 gaccgatcca gcctccggac tctagcgttt aaacgctagc caccatggcg tacccatacg 4321 atgttccaga ttacgctgct cgaggttatc aaacaagttt gtacaaaaaa gttggcatgg 4381 ccgtcatggc gccccgaacc ctcgtcctgc tactctcggg ggccctggcc ctgacccaga 4441 cctgggcggg ctcccactcc atgaggtatt tctacacctc cgtgtcccgg cccggccgcg 4501 gggagccccg cttcatcgcc gtgggctacg tggacgacac gcagttcgtg cggttcgaca 4561 gcgacgccgc gagccagagg atggagccgc gggcgccgtg gatagagcag gaggggccgg 4621 agtattggga ccggaacaca cggaatgtga aggcccactc acagactgac cgagagagcc 4681 tgcggatcgc gctccgctac tacaaccaga gcgaggacgg ttctcacacc atccagagga 4741 tgtatggctg cgacgtgggg ccggacgggc gcttcctccg cgggtaccag caggacgctt 4801 acgacggcaa ggattacatc gccctgaacg aggacctgcg ctcttggacc gcggcggaca 4861 tggcggctca gatcacccag cgcaagtggg agacggccca tgaggcggag cagtggagag 4921 cctacctgga gggccggtgc gtggagtggc tccgcagata cctggagaac gggaaggaga 4981 cgctgcagcg cacggacgcc cccaagacgc atatgactca ccacgctgtc tctgaccatg 5041 aggccaccct gaggtgctgg gccctgagct tctaccctgc ggagatcaca ctgacctggc 5101 agcgggatgg ggaggaccag acccaggaca cggagctcgt ggagaccagg cctgcagggg 5161 atgggacctt ccagaagtgg gcgtctgtgg tggtgccttc tggacaggag cagagataca 5221 cctgccatgt gcagcatgag ggtctgccca agcccctcac cctgagatgg gagccgtctt 5281 cccagcccac catccccatc gtgggcatca ttgctggcct ggttctcttt ggagctgtga 5341 tcgctggagc tgtggtcgct gctgtgatgt ggaggaggaa gagctcagat agaaaaggag 5401 ggagctactc tcaggctgca agcagtgaca gtgcccaggg ctctgatatg tctctcacag 5461 cttgtaaagt gtgcccaact ttcttgtaca aagtggttcg ataacgaatt ctaccgggta 5521 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact 5581 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa 5641 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag 5701 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc 5761 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc 5821 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg 5881 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga 5941 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca 6001 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc 6061 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg 6121 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc 6181 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg 6241 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg 6301 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga 6361 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg 6421 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg 6481 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc 6541 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc 6601 gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc 6661 cgaccgaaag gagcgcacga ccccatgcat cgatagatcc taatcaacct ctggattaca 6721 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 6781 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 6841 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 6901 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 6961 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 7021 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 7081 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga 7141 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 7201 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 7261 gtcggatctc cctttgggcc gcctccccgc ctgagatcct ttaagaccaa tgacttacaa 7321 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca 7381 ctcccaacga agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat 7441 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt 7501 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc 7561 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat 7621 tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc ccgggttaat 7681 taaggaaagg gctagatcat tcttgaagac gaaagggcct cgtgatacgc ctatttttat 7741 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg 7801 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga 7861 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac 7921 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc 7981 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca 8041 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc 8101 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg 8161 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac 8221 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca 8281 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg 8341 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac 8401 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg 8461 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat 8521 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg 8581 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg 8641 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc 8701 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc 8761 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt 8821 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt 8881 aacgtgagtt ttcgttccac tgagcgtca //