LOCUS Exported 6255 bp ds-DNA circular SYN 21-JUL-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV Fcgamma RIIa IRES neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6255) AUTHORS Trivedi V, Zhang SC, Castoreno AB, Stockinger W, Shieh EC, Vyas JM, Frickel EM, Nohturfft A TITLE Immunoglobulin G signaling activates lysosome/phagosome docking. JOURNAL Proc Natl Acad Sci U S A. 2006 Nov 28. 103(48):18226-31. PUBMED 17110435 REFERENCE 2 (bases 1 to 6255) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6255 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 381..584 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 535..555 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 629..648 /label=T7 /note="T7 promoter, forward primer" promoter 629..647 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" intron 1762..1991 /label=chimeric intron /note="chimera between introns from adenovirus and immunoglobulin heavy chain genes" primer_bind 1974..1993 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 2064..2637 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2231..2248) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2458..2477 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2673..3476 /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2736..2755) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3346..3365 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(3508..3525) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3514..3738 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind 3785..3804 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3864..3880) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3864..3880) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3877..3899) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3888..3904 /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(3912..3942) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3957..3978 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4095..4112) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4266..4854) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4346..4365) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5025..5885) /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 5648..5667 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5886..5990) /gene="bla" /label=AmpR promoter primer_bind complement(6065..6084) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc 421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt 481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa 541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag 601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt 661 ggtaccgagc tcggatcgat atctgcggcc tagctagcgc ttaaggcctg ttaaccggtc 721 gtacgtctcc ggattcgaat tcaccatgga gacccaaatg tctcagaatg tatgtcccag 781 aaacctgtgg ctgcttcaac cattgacagt tttgctgctg ctggcttctg cagacagtca 841 agctgcagct cccccaaagg ctgtgctgaa acttgagccc ccgtggatca acgtgctcca 901 ggaggactct gtgactctga catgccaggg ggctcgcagc cctgagagcg actccattca 961 gtggttccac aatgggaatc tcattcccac ccacacgcag cccagctaca ggttcaaggc 1021 caacaacaat gacagcgggg agtacacgtg ccagactggc cagaccagcc tcagcgaccc 1081 tgtgcatctg actgtgcttt ccgaatggct ggtgctccag acccctcacc tggagttcca 1141 ggagggagaa accatcatgc tgaggtgcca cagctggaag gacaagcctc tggtcaaggt 1201 cacattcttc cagaatggaa aatcccagaa attctcccgt ttggatccca ccttctccat 1261 cccacaagca aaccacagtc acagtggtga ttaccactgc acaggaaaca taggctacac 1321 gctgttctca tccaagcctg tgaccatcac tgtccaagtg cccagcatgg gcagctcttc 1381 accaatgggg atcattgtgg ctgtggtcat tgcgactgct gtagcagcca ttgttgctgc 1441 tgtagtggcc ttgatctact gcaggaaaaa gcggatttca gccaattcca ctgatcctgt 1501 gaaggctgcc caatttgagc cacctggacg tcaaatgatt gccatcagaa agagacaact 1561 tgaagaaacc aacaatgact atgaaacagc tgacggcggc tacatgactc tgaaccccag 1621 ggcacctact gacgatgata aaaacatcta cctgactctt cctcccaacg accatgtcaa 1681 cagtaataac taagcggccg catagataac tgatccagtg tgctggaatt aattcgctgt 1741 ctgcgagggc cagctgttgg ggtgagtact ccctctcaaa agcgggcatg acttctgcgc 1801 taagattgtc agtttccaaa aacgaggagg atttgatatt cacctggccc gcggtgatgc 1861 ctttgagggt ggccgcgtcc atctggtcag aaaagacaat ctttttgttg tcaagcttga 1921 ggtgtggcag gcttgagatc tggccataca cttgagtgac aatgacatcc actttgcctt 1981 tctctccaca ggtgtccact cccaggtcca actgcaggtc gagcatgcat ctagggcggc 2041 caattccgcc cctctccccc ccccccctct ccctcccccc cccctaacgt tactggccga 2101 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg 2161 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg 2221 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt 2281 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac 2341 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca 2401 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg 2461 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg 2521 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa 2581 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataagct 2641 tgccacaacc cgggataatt cctgcagcca atatgggatc ggccattgaa caagatggat 2701 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 2761 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 2821 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc 2881 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 2941 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 3001 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 3061 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 3121 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 3181 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgatgat ctcgtcgtga 3241 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 3301 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 3361 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 3421 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagggg 3481 atcaattctc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg 3541 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 3601 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 3661 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 3721 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg agtgcattct 3781 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct 3841 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 3901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 3961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 4021 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 4081 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 4141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 4201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 4681 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca 4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 4921 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 4981 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 5041 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 5101 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 5161 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 5221 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 5281 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 5341 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 5401 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 5461 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 5521 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 5581 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 5641 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 5701 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 5761 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 5821 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 5881 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 5941 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6001 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc 6061 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt 6121 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg 6181 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg 6241 ccagatatac gcgtt //