LOCUS V035145 6655 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035145 VERSION V035145 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 964..2145 /label="NDRG1(NM_006096)" /note="NDRG1(NM_006096)" /gene="NDRG1" CDS 2152..2217 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2255..2479 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2525..2953 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2967..3296 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3363..4157 /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 4331..4452 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4501..4517) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4525..4541 /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(4549..4579) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4594..4615 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4903..5488) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5659..6519) /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(6520..6624) /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 atccactagt ccagtgtggt ggaattcgcc 961 accatgtctc gggagatgca ggatgtagac ctcgctgagg tgaagccttt ggtggagaaa 1021 ggggagacca tcaccggcct cctgcaagag tttgatgtcc aggagcagga catcgagact 1081 ttacatggct ctgttcacgt cacgctgtgt gggactccca agggaaaccg gcctgtcatc 1141 ctcacctacc atgacatcgg catgaaccac aaaacctgct acaaccccct cttcaactac 1201 gaggacatgc aggagatcac ccagcacttt gccgtctgcc acgtggacgc ccctggccag 1261 caggacggcg cagcctcctt ccccgcaggg tacatgtacc cctccatgga tcagctggct 1321 gaaatgcttc ctggagtcct tcaacagttt gggctgaaaa gcattattgg catgggaaca 1381 ggagcaggcg cctacatcct aactcgattt gctctaaaca accctgagat ggtggagggc 1441 cttgtcctta tcaacgtgaa cccttgtgcg gaaggctgga tggactgggc cgcctccaag 1501 atctcaggat ggacccaagc tctgccggac atggtggtgt cccacctttt tgggaaggaa 1561 gaaatgcaga gtaacgtgga agtggtccac acctaccgcc agcacattgt gaatgacatg 1621 aaccccggca acctgcacct gttcatcaat gcctacaaca gccggcgcga cctggagatt 1681 gagcgaccaa tgccgggaac ccacacagtc accctgcagt gccctgctct gttggtggtt 1741 ggggacagct cgcctgcagt ggatgccgtg gtggagtgca actcaaaatt ggacccaaca 1801 aagaccactc tcctcaagat ggcggactgt ggcggcctcc cgcagatctc ccagccggcc 1861 aagctcgctg aggccttcaa gtacttcgtg cagggcatgg gatacatgcc ctcggctagc 1921 atgacccgcc tgatgcggtc ccgcacagcc tctggttcca gcgtcacttc tctggatggc 1981 acccgcagcc gctcccacac cagcgagggc acccgaagcc gctcccacac cagcgagggc 2041 acccgcagcc gctcgcacac cagcgagggg gcccacctgg acatcacccc caactcgggt 2101 gctgctggga acagcgccgg gcccaagtcc atggaggtct cctgcctcga ggactacaaa 2161 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag 2221 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct 2281 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt 2341 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg 2401 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg 2461 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat 2521 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 2581 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 2641 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 2701 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt 2761 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 2821 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat 2881 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 2941 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct 3001 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa 3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3121 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt 3181 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct 3241 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc 3301 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc 3361 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat 3421 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt 3481 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac 3541 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg 3601 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc 3661 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa 3721 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc 3781 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg 3841 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg 3901 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa 3961 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg 4021 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct 4081 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc 4141 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa 4201 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat 4261 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt 4321 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 4381 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 4441 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg 4501 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4561 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4621 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4681 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4741 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4801 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4861 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4921 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4981 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 5041 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 5101 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 5161 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 5221 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 5281 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5341 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5401 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca 5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //