LOCUS V023185 6294 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V023185 VERSION V023185 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers promoter 123..1301 /note="strong constitutive promoter for human elongation factor EF-1α" /label="EF-1α promoter" CDS 1364..2992 /label="Pfkfb3(NM_001177756)" /note="Pfkfb3(NM_001177756)" /gene="Pfkfb3" CDS 2993..3010 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 3135..3256 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" rep_origin complement(3263..3718) /direction=LEFT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3745..3849 /gene="bla" /label="AmpR promoter" promoter 3851..4208 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4243..5037 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5269..5316 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" rep_origin 5645..6233 /direction=RIGHT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" ORIGIN 1 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt 61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa 121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg 181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa 241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg 301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg 361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa 421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt 481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct 541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg 601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct 661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg 721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc 781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc 841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac 901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga 961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc 1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg 1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga 1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt 1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga 1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtacgct 1321 agcgtttaaa cttaagcttg gtaccgagct cggatccgcc accatgccgt tggaactgac 1381 ccagagccgg gtacagaaga tctgggtgcc cgtcgatcac cgaccctcgt tgcccagatc 1441 ctgtgggccg aagctgacca actccccaac cgtgattgtg atggtggggc tcccagcccg 1501 gggtaagact tacatctcca agaagctgac tcgctacctc aactggatag gtgttccaac 1561 gaaagtgttc aatgtgggag agtatcggcg tgaggctgtg aagcagtaca gctcttacaa 1621 cttcttccgc cctgacaatg aggaagccat gagagtccga aaacagtgtg ccttagctgc 1681 cctgagagat gtcaaaagct acctgacaaa ggaaggtgga cagattgcag tttttgatgc 1741 caccaataca actagagaaa gaaggcacat gatccttaat tttgccaaag aaaatgactt 1801 caaggcattt ttcatcgagt cggtctgtga cgaccctact gttgtggcct ccaacatcat 1861 ggaagttaaa atctccagcc cggattacaa agactgcaac tcagcagaag ccatggacga 1921 cttcatgaag agaatcaatt gctatgaagc cagctaccag cctcttgacc ctgataaatg 1981 tgacagggac ttgtcactca tcaaagtgat tgatgttggc cggaggttct tggtgaacag 2041 agtgcaggac cacatccaga gccgaattgt atactacctg atgaacatcc acgtgcagcc 2101 ccgtaccatc tacctgtgtc ggcatggcga gaatgagtac aacttgcagg gcaagattgg 2161 gggcgactca ggcttgtcca gcagaggcaa gaagttcgcc aatgccctga gcaagttcgt 2221 agaagaacag aatctgaagg acctccgtgt gtggaccagc cagctgaaaa gtaccattca 2281 gaccgcggag gccctgaggc tcccctacga gcagtggaag gcactcaacg agatcgatgc 2341 tggtgtgtgt gaggaactaa cctatgagga gatcagagat acctaccctg aggagtacgc 2401 actgcgtgaa caggacaagt actactatcg ctatcccacg ggagagtcct accaggatct 2461 tgtccaacgc ctggagcctg tgatcatgga attagagcgc caagagaatg tattggtcat 2521 ctgtcaccag gctgttctac gctgcctact agcctacttc ctcgacaaga gtgcagagga 2581 gatgccatac ctgaaatgtc cgctccacac tgtcctgaaa ctgacacctg tggcatatgg 2641 gtgccgggtg gagtccattt acttgaatgt agaatcggtg agcacacacc gggagaggtc 2701 agaggctgtc aaaatacaac actttgccag tgtagtcaga ccttcctctt acaccgaact 2761 tgactttcag tctgtggagt ccgcaaaaca ggatgcaaag aagggaccta acccgctcat 2821 gagacgcaat agtgtcaccc cactagccag tcccgagccc accaaaaagc ctcgcatcaa 2881 cagctttgag gagcgtgtgg cttccacttc tgctgccctg ccaagctgcc tgcccccgga 2941 agtgcccacg cagctgcccg gacaaccttt gctagggaaa gcctgtctaa cacatcatca 3001 ccatcaccat tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg 3061 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg 3121 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 3181 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 3241 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta 3301 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 3361 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 3421 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 3481 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 3541 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 3601 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 3661 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca 3721 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 3781 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 3841 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa 3901 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3961 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 4021 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4081 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4141 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4201 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 4261 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 4321 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 4381 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg 4441 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4501 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4561 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4621 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4681 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4741 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg 4801 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4861 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4921 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4981 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 5041 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 5101 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 5161 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc 5221 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa 5281 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg 5341 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt 5401 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg 5461 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac 5521 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 5581 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 5641 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 5701 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 5761 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc 5821 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 5881 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 5941 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 6001 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 6061 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 6121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 6181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 6241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgt //