LOCUS V025966 5924 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025966 VERSION V025966 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..1414 /label="REG4(NM_032044)" /note="REG4(NM_032044)" /gene="REG4" CDS 1421..1486 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1524..1748 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1794..2222 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2236..2565 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2632..3426 /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 3600..3721 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3770..3786) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3794..3810 /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(3818..3848) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3863..3884 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4172..4757) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4928..5788) /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(5789..5893) /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 atggcttcca gaagcatgcg 961 gctgctccta ttgctgagct gcctggccaa aacaggagtc ctgggtgata tcatcatgag 1021 acccagctgt gctcctggat ggttttacca caagtccaat tgctatggtt acttcaggaa 1081 gctgaggaac tggtctgatg ccgagctcga gtgtcagtct tacggaaacg gagcccacct 1141 ggcatctatc ctgagtttaa aggaagccag caccatagca gagtacataa gtggctatca 1201 gagaagccag ccgatatgga ttggcctgca cgacccacag aagaggcagc agtggcagtg 1261 gattgatggg gccatgtatc tgtacagatc ctggtctggc aagtccatgg gtgggaacaa 1321 gcactgtgct gagatgagct ccaataacaa ctttttaact tggagcagca acgaatgcaa 1381 caagcgccaa cacttcctgt gcaagtaccg accactcgag gactacaaag accatgacgg 1441 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt 1501 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc 1561 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa 1621 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg 1681 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg 1741 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc 1801 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 1861 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 1921 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 1981 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 2041 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 2101 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 2161 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 2221 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc 2281 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc 2341 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg 2401 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat 2461 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc 2521 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct 2581 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa 2641 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac 2701 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg 2761 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag 2821 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt 2881 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg 2941 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg 3001 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga 3061 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag 3121 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat 3181 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt 3241 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg 3301 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt 3361 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc 3421 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac 3481 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg 3541 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca 3601 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 3661 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 3721 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt 3781 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 3841 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 3901 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 3961 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 4021 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 4081 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 4141 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 4201 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 4261 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 4321 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 4381 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 4441 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 4501 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 4561 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 4621 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4681 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca 4741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga 4801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga 4861 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt 4921 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt 4981 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat 5041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag 5101 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct 5161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt 5221 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg 5281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca 5341 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt 5401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat 5461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac 5521 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa 5581 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt 5641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt 5701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa 5761 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt 5821 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa 5881 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc //