LOCUS Exported 6575 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses a fluorescent protein, mVenus. ACCESSION . VERSION . KEYWORDS pEF.DEST51-mVenus SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6575) TITLE A feed-forward loop between SorLA and HER3 determines heregulin response and neratinib resistance JOURNAL Unpublished REFERENCE 2 (bases 1 to 6575) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6575 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(166..185) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 473..1651 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 704..1642 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1599..1619 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" primer_bind 1668..1687 /label=T7 /note="T7 promoter, forward primer" promoter 1668..1686 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1718..1742 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 1780..1789 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1786..2505 /codon_start=1 /product="Venus YFP with monomerizing A206K mutation (Nagai et al., 2002; Kremers et al., 2006)" /label=mVenus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1831..1852) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2092..2111) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2439..2460 /label=EGFP-C /note="EGFP, forward primer" protein_bind complement(2523..2547) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2573..2614 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 2624..2641 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2664..2681) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2670..2894 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2940..3368 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3027..3046) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3237..3258 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3377..3397) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3382..3711 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3562..3697 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3624..3643 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3759..3806 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3825..4223 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 4381..4502 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4418..4437) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4472..4491 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4551..4567) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4551..4567) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4564..4586) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4575..4591 /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(4599..4629) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4644..4665 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4782..4799) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4942..5530) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5022..5041) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5701..6561) /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 6324..6343 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6562..91) /gene="bla" /label=AmpR promoter ORIGIN 1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga 421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg 481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt 541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt 601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc 661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg 721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac 781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc 841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg 901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa 961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag 1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg 1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc 1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc 1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg 1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg 1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc 1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc 1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc 1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt 1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca 1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac 1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg 1741 ctggcgccgg aaccaattca gtcgactgga tccggtaccg ccaccatggt gagcaagggc 1801 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 1861 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 1921 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 1981 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 2041 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 2101 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 2161 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 2221 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac 2281 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag 2341 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag 2401 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 2461 accgccgccg ggatcactct cggcatggac gagctgtaca agtaatgact cgagatatct 2521 agacccagct ttcttgtaca aagtggttga tctagagggc ccgcggttcg aaggtaagcc 2581 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca 2641 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 2701 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 2761 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 2821 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 2881 ggtgggcgct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca 2941 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 3001 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 3061 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 3121 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 3181 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 3241 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 3301 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 3361 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca 3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc 3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata 3541 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg 3601 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag 3661 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg 3721 ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa tcatcggcat 3781 agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc aagcctttgt 3841 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct 3901 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg 3961 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg 4021 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca 4081 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag 4141 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct 4201 ctggttatgt gtgggagggc taagcacttc gtggccgagg agcaggactg acacgtgcta 4261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4321 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4501 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4561 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4621 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4681 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4741 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4801 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca ggaaccgtaa 4921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5341 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5461 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5761 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 5821 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 5881 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 5941 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6001 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6061 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6121 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6181 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6241 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6301 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6361 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6421 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6481 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6541 acggaaatgt tgaatactca tactcttcct ttttc //