LOCUS V040728 6877 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040728 VERSION V040728 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..2367 /label="FOXN3(NM_005197)" /note="FOXN3(NM_005197)" /gene="FOXN3" CDS 2374..2439 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2477..2701 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2747..3175 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3189..3518 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3585..4379 /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 4553..4674 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4723..4739) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4747..4763 /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(4771..4801) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4816..4837 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5125..5710) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5881..6741) /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(6742..6846) /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 accatgggtc cagtcatgcc tcccagtaag aagccagaaa gctcaggaat tagtgtctcc 1021 agtggactga gtcagtgtta cgggggcagc ggtttctcca aggcccttca ggaagacgat 1081 gacctcgact tttctctgcc tgacatccga ttagaagagg gggccatgga agatgaagag 1141 ctgaccaacc tgaactggct gcacgagagc aagaacttgc tgaagagctt tggggagtcg 1201 gtcctcagga gtgtcagccc cgtccaggac ctggacgatg acaccccccc atcccctgcc 1261 cactctgaca tgccctacga tgccaggcag aaccccaact gcaaaccccc ctactccttc 1321 agctgcctca tatttatggc catcgaggac tctccaacca agcgcctgcc agtgaaggat 1381 atctacaact ggatcttgga acattttccg tattttgcaa atgcacctac tgggtggaaa 1441 aactcagtga gacacaattt atcattgaat aagtgtttta agaaagtgga caaagagagg 1501 agtcagagta ttgggaaagg gtcgttgtgg tgcatagacc cagagtatag acaaaatcta 1561 attcaggctt tgaaaaagac accttatcac ccacacccac acgtgttcaa tacacctccc 1621 acctgtcctc aggcatatca aagcacatca ggtccaccca tctggccggg cagtaccttc 1681 ttcaagagaa atggagccct tctccaagtt cctccaggag tgatccaaaa tggagcgcgg 1741 gtcctgagcc gagggctgtt tcctggcgtg cggccgctgc caatcactcc cattggggtg 1801 acagcggcca tgaggaatgg catcaccagc tgccggatgc ggactgagag tgagccatct 1861 tgtggctccc cagtggtcag cggagacccc aaggaggatc acaactacag cagtgccaag 1921 tcctccaacg cccggagcac ctcgcccacc agcgactcca tctcctcctc ctcctcctca 1981 gccgacgacc actatgagtt tgccaccaag gggagccagg agggcagcga gggcagcgag 2041 gggagcttcc ggagccacga gagccccagc gacacggaag aggacgacag gaagcacagc 2101 cagaaggagc ccaaggattc tctgggggac agcgggtacg catcccagca caagaagcgc 2161 cagcacttcg ccaaggccag gaaggtcccc agcgacacac tgcccctcaa aaagagacgc 2221 accgaaaagc cccccgagag cgatgatgag gagatgaaag aagcggcagg gtccctcctg 2281 cacttagcag ggatccggtc ctgtttgaat aacatcacca atcggacggc aaaggggcag 2341 aaagagcaaa aggaaaccac aaaaaatctc gaggactaca aagaccatga cggtgattat 2401 aaagatcatg acatcgacta caaggatgac gatgacaagt agtgagggcc cgtttaaacc 2461 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2521 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2581 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2641 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2701 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc 2761 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc 2821 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt 2881 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac 2941 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 3001 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 3061 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 3121 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc 3181 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta 3241 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag 3301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa 3361 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac 3421 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt 3481 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat 3541 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg 3601 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac 3661 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg 3721 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc 3781 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg 3841 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc 3901 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc 3961 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta 4021 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg 4081 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg 4141 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat 4201 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc 4261 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta 4321 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag 4381 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt 4441 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg 4501 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt 4561 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4621 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt 4681 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt 4741 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa 4801 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc 4861 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag 4921 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt 4981 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga 5041 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg 5101 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa 5161 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt 5221 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct 5281 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct 5341 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc 5401 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt 5461 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc 5521 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat 5581 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa 5641 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5701 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5761 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 5821 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 5881 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 5941 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 6001 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 6061 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 6121 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 6181 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 6241 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 6301 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 6361 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 6421 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 6481 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 6541 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 6601 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 6661 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 6721 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 6781 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 6841 tccgcgcaca tttccccgaa aagtgccacc tgacgtc //