LOCUS V025355 6002 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025355 VERSION V025355 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 941..1492 /label="GREM1(NM_013372)" /note="GREM1(NM_013372)" /gene="GREM1" CDS 1499..1564 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1602..1826 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1872..2300 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2314..2643 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2710..3504 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 3678..3799 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3848..3864) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3872..3888 /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(3896..3926) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3941..3962 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4250..4835) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5006..5866) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(5867..5971) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgagccgca cagcctacac 961 ggtgggagcc ctgcttctcc tcttggggac cctgctgccg gctgctgaag ggaaaaagaa 1021 agggtcccaa ggtgccatcc ccccgccaga caaggcccag cacaatgact cagagcagac 1081 tcagtcgccc cagcagcctg gctccaggaa ccgggggcgg ggccaagggc ggggcactgc 1141 catgcccggg gaggaggtgc tggagtccag ccaagaggcc ctgcatgtga cggagcgcaa 1201 atacctgaag cgagactggt gcaaaaccca gccgcttaag cagaccatcc acgaggaagg 1261 ctgcaacagt cgcaccatca tcaaccgctt ctgttacggc cagtgcaact ctttctacat 1321 ccccaggcac atccggaagg aggaaggttc ctttcagtcc tgctccttct gcaagcccaa 1381 gaaattcact accatgatgg tcacactcaa ctgccctgaa ctacagccac ctaccaagaa 1441 gaagagagtc acacgtgtga agcagtgtcg ttgcatatcc atcgatttgg atctcgagga 1501 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga 1561 caagtagtga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca 1621 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 1681 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 1741 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 1801 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 1861 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 1921 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 1981 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 2041 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 2101 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 2161 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 2221 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 2281 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 2341 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 2401 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 2461 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 2521 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 2581 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2641 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 2701 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 2761 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 2821 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 2881 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 2941 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 3001 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 3061 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 3121 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 3181 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 3241 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 3301 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 3361 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 3421 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 3481 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 3541 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 3601 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 3661 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 3721 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 3781 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 3841 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 3901 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 3961 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 4021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 4081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 4141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 4201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 4261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 4321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 4681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca 4801 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 4861 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 4921 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 4981 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 5041 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 5101 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 5161 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 5221 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 5281 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 5341 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 5401 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 5461 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 5521 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 5581 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 5641 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 5701 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 5761 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 5821 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 5881 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 5941 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 6001 tc //