LOCUS V041022 6155 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V041022 VERSION V041022 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 907..1602 /label="DDIT4(NM_019058)" /note="DDIT4(NM_019058)" /gene="DDIT4" CDS 1612..1653 /codon_start=1 /product="epitope tag from simian virus 5" /transl_table=1 /translation="GKPIPNPLLGLDST" /label="V5 tag" polyA_signal 1755..1979 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2025..2453 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2467..2796 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2863..3657 /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 3831..3952 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4001..4017) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4025..4041 /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(4049..4079) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4094..4115 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4403..4988) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5159..6019) /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(6020..6124) /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 gccaccatgc ctagcctttg ggaccgcttc tcgtcgtcgt ccacctcctc ttcgccctcg 961 tccttgcccc gaactcccac cccagatcgg ccgccgcgct cagcctgggg gtcggcgacc 1021 cgggaggagg ggtttgaccg ctccacgagc ctggagagct cggactgcga gtccctggac 1081 agcagcaaca gtggcttcgg gccggaggaa gacacggctt acctggatgg ggtgtcgttg 1141 cccgacttcg agctgctcag tgaccctgag gatgaacact tgtgtgccaa cctgatgcag 1201 ctgctgcagg agagcctggc ccaggcgcgg ctgggctctc gacgccctgc gcgcctgctg 1261 atgcctagcc agttggtaag ccaggtgggc aaagaactac tgcgcctggc ctacagcgag 1321 ccgtgcggcc tgcggggggc gctgctggac gtctgcgtgg agcagggcaa gagctgccac 1381 agcgtgggcc agctggcact cgaccccagc ctggtgccca ccttccagct gaccctcgtg 1441 ctgcgcctgg actcacgact ctggcccaag atccaggggc tgtttagctc cgccaactct 1501 cccttcctcc ctggcttcag ccagtccctg acgctgagca ctggcttccg agtcatcaag 1561 aagaagctgt acagctcgga acagctgctc attgaggagt gtaagcttat gggtaagcct 1621 atccctaacc ctctcctcgg tctcgattct acataggatc cactagtcca gtgtggtgga 1681 attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg tttaaacccg 1741 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 1801 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 1861 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 1921 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 1981 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2041 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 2101 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2161 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2221 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2281 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2341 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2401 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2461 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2521 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2581 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 2641 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 2701 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 2761 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 2821 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 2881 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 2941 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3001 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3061 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3121 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3181 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3241 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3301 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3361 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3421 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3481 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3541 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 3601 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 3661 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 3721 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 3781 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 3841 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 3901 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 3961 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4021 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4081 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4141 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4201 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4261 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4321 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4381 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4441 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4501 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4561 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4621 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4681 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4741 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4801 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4861 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4921 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4981 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5041 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 5101 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5161 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5221 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5281 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5341 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5401 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5461 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5521 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5581 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 5641 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 5701 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 5761 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 5821 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 5881 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 5941 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6001 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6061 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6121 cgcgcacatt tccccgaaaa gtgccacctg acgtc //