LOCUS Exported 7459 bp ds-DNA circular SYN 19-APR-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7459) AUTHORS Trial User TITLE Direct Submission JOURNAL Exported Apr 19, 2018 from SnapGene 4.1.4 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7459 /organism="recombinant plasmid" /mol_type="other DNA" enhancer 93..472 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 473..676 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 721..739 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 762..786 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 788..797 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 790..2040 /codon_start=1 /product="human lysosome-associated membrane glycoprotein 1" /label=LAMP1 /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV LIAYLVGRKRSHAGYQTI" protein_bind complement(2042..2066) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(2082..2102) /label=PLAMP1-pmturquoise-BB.REV primer_bind 2085..2106 /label=plamp1-PMTURQUOISE-INS.FOR CDS 2086..2802 /codon_start=1 /product="enhanced monomeric variant of CFP (Goedhart et al., 2012)" /label=mTurquoise2 /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" regulatory 2086..2089 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(2776..2797) /label=plamp1-PMTURQUOISE-INS.REV primer_bind 2782..2807 /label=PLAMP1-pmturquoise-BB.FOR polyA_signal 2914..3138 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3184..3612 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3626..3955 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3806..3941 /label=SV40 ori /note="SV40 origin of replication" CDS 4022..4816 /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" polyA_signal 4990..5111 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5160..5176) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 5184..5200 /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(5208..5238) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5253..5274 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5562..6150) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6321..7181) /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" promoter complement(7182..7286) /gene="bla" /label=AmpR promoter ORIGIN 1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct 61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata 121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg 481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct 541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa 601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt 661 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat 721 taatacgact cactataggg agacccaagc tggctagatc aacaagtttg tacaaaaaag 781 caggctacca tggcggcccc cggcagcgcc cggcgacccc tgctgctgct actgctgttg 841 ctgctgctcg gcctcatgca ttgtgcgtca gcagcaatgt ttatggtgaa aaatggcaac 901 gggaccgcgt gcataatggc caacttctct gctgccttct cagtgaacta cgacaccaag 961 agtggcccta agaacatgac ctttgacctg ccatcagatg ccacagtggt gctcaaccgc 1021 agctcctgtg gaaaagagaa cacttctgac cccagtctcg tgattgcttt tggaagagga 1081 catacactca ctctcaattt cacgagaaat gcaacacgtt acagcgtcca gctcatgagt 1141 tttgtttata acttgtcaga cacacacctt ttccccaatg cgagctccaa agaaatcaag 1201 actgtggaat ctataactga catcagggca gatatagata aaaaatacag atgtgttagt 1261 ggcacccagg tccacatgaa caacgtgacc gtaacgctcc atgatgccac catccaggcg 1321 tacctttcca acagcagctt cagcagggga gagacacgct gtgaacaaga caggccttcc 1381 ccaaccacag cgccccctgc gccacccagc ccctcgccct cacccgtgcc caagagcccc 1441 tctgtggaca agtacaacgt gagcggcacc aacgggacct gcctgctggc cagcatgggg 1501 ctgcagctga acctcaccta tgagaggaag gacaacacga cggtgacaag gcttctcaac 1561 atcaacccca acaagacctc ggccagcggg agctgcggcg cccacctggt gactctggag 1621 ctgcacagcg agggcaccac cgtcctgctc ttccagttcg ggatgaatgc aagttctagc 1681 cggtttttcc tacaaggaat ccagttgaat acaattcttc ctgacgccag agaccctgcc 1741 tttaaagctg ccaacggctc cctgcgagcg ctgcaggcca cagtcggcaa ttcctacaag 1801 tgcaacgcgg aggagcacgt ccgtgtcacg aaggcgtttt cagtcaatat attcaaagtg 1861 tgggtccagg ctttcaaggt ggaaggtggc cagtttggct ctgtggagga gtgtctgctg 1921 gacgagaaca gcatgctgat ccccatcgct gtgggtggtg ccctggcggg gctggtcctc 1981 atcgtcctca tcgcctacct cgtcggcagg aagaggagtc acgcaggcta ccagactatc 2041 cacccagctt tcttgtacaa agtggttgat ctagcgttta ccggtatggt gagcaagggc 2101 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc 2161 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg 2221 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg 2281 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc 2341 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc 2401 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag 2461 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac 2521 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac 2581 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag 2641 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag 2701 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg 2761 accgccgccg ggatcactct cggcatggac gagctgtaca agggtaccga gctcggatcc 2821 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta 2881 gagggcccgt tgaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg 2941 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3001 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3061 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3121 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc 3181 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 3241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 3301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 3361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 3421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 3481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 3541 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 3601 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc 3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 3961 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg 4021 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 4081 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 4141 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 4201 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 4261 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 4321 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 4381 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 4441 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 4501 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 4561 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 4621 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 4681 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 4741 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 4801 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 4861 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 5041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 5101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 5161 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 5221 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 5281 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 5341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 5401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 5461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 5761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 5821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 6001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 6061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 6121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 6181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 6241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 6301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 6361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 6421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 6481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 6541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 7021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 7081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 7141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 7201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 7261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac 7321 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg 7381 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga 7441 gcaaaattta agctacaac //