LOCUS V033562 6454 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V033562 VERSION V033562 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 925..2205 /label="KCNJ2(NM_000891)" /note="KCNJ2(NM_000891)" /gene="KCNJ2" CDS 2218..2283 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2318..2542 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2588..3016 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3030..3359 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3371..3970 /codon_start=1 /gene="pac from Streptomyces alboniger" /note="confers resistance to puromycin" /product="puromycin N-acetyltransferase" /transl_table=1 /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*" /label="PuroR" polyA_signal 4130..4251 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4300..4316) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4324..4340 /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(4348..4378) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4393..4414 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4702..5287) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5458..6318) /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(6319..6423) /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 aagctttcta gagaattcgc caccatgggc agtgtgcgaa ccaaccgcta cagcatcgtc 961 tcttcagaag aagacggtat gaagttggcc accatggcag ttgcaaatgg ctttgggaac 1021 gggaagagta aagtccacac ccgacaacag tgcaggagcc gctttgtgaa gaaagatggc 1081 cactgtaatg ttcagttcat caatgtgggt gagaaggggc aacggtacct cgcagacatc 1141 ttcaccacgt gtgtggacat tcgctggcgg tggatgctgg ttatcttctg cctggctttc 1201 gtcctgtcat ggctgttttt tggctgtgtg ttttggttga tagctctgct ccatggggac 1261 ctggatgcat ccaaagaggg caaagcttgt gtgtccgagg tcaacagctt cacggctgcc 1321 ttcctcttct ccattgagac ccagacaacc ataggctatg gtttcagatg tgtcacggat 1381 gaatgcccaa ttgctgtttt catggtggtg ttccagtcaa tcgtgggctg catcatcgat 1441 gctttcatca ttggcgcagt catggccaag atggcaaagc caaagaagag aaacgagact 1501 cttgtcttca gtcacaatgc cgtgattgcc atgagagacg gcaagctgtg tttgatgtgg 1561 cgagtgggca atcttcggaa aagccacttg gtggaagctc atgttcgagc acagctcctc 1621 aaatccagaa ttacttctga aggggagtat atccctctgg atcaaataga catcaatgtt 1681 gggtttgaca gtggaatcga tcgtatattt ctggtgtccc caatcactat agtccatgaa 1741 atagatgaag acagtccttt atatgatttg agtaaacagg acattgacaa cgcagacttt 1801 gaaatcgtgg tcatactgga aggcatggtg gaagccactg ccatgacgac acagtgccgt 1861 agctcttatc tagcaaatga aatcctgtgg ggccaccgct atgagcctgt gctctttgaa 1921 gagaagcact actacaaagt ggactattcc aggttccaca aaacttacga agtccccaac 1981 actccccttt gtagtgccag agacttagca gaaaagaaat atatcctctc aaatgcaaat 2041 tcattttgct atgaaaatga agttgccctc acaagcaaag aggaagacga cagtgaaaat 2101 ggagttccag aaagcactag tacggacacg ccccctgaca tagaccttca caaccaggca 2161 agtgtacctc tagagcccag gcccttacgg cgagagtcgg agatactcga gaccggtgac 2221 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac 2281 aagtgaccgg taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca 2341 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc 2401 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg 2461 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct 2521 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg 2581 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 2641 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 2701 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc 2761 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 2821 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 2881 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 2941 gatttataag ggattttggc catttcggcc tattggttaa aaaatgagct gatttaacaa 3001 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 3061 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 3181 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 3241 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 3301 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 3361 agctcccggg atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc 3421 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt 3481 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt 3541 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac 3601 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga 3661 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg 3721 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa 3781 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc 3841 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac 3901 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc 3961 cggtgcctga ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 4021 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4081 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat 4141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt 4201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg 4261 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 4441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 4501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 4561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 4621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 4681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5221 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5281 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 5401 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 5461 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 5521 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 5581 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5641 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5701 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 5761 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 5821 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 5881 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 5941 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 6001 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 6061 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 6121 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6181 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6241 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6301 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 6421 gcgcacattt ccccgaaaag tgccacctga cgtc //