LOCUS V031094 6087 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031094 VERSION V031094 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers primer_bind 166..182 /note="common sequencing primer, one of multiple similar variants" /label="M13 fwd" enhancer 343..722 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 723..926 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 952..970 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 1029..2234 /label="Lhx5(NM_008499)" /note="Lhx5(NM_008499)" /gene="Lhx5" CDS 2253..2282 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /transl_table=1 /translation="EQKLISEEDL" /label="Myc" CDS 2301..2324 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" primer_bind complement(2344..2360) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" polyA_signal 2371..2993 /note="human growth hormone polyadenylation signal" /label="hGH poly(A) signal" rep_origin complement(3142..3730) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" polyA_signal 4059..4106 /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" /label="HSV TK poly(A) signal" CDS complement(4338..5132) /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" promoter complement(5167..5524) /note="SV40 enhancer and early promoter" /label="SV40 promoter" promoter complement(5526..5630) /gene="bla" /label="AmpR promoter" rep_origin 5657..6085 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" ORIGIN 1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag 61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt 241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata 301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta 361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg 421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga 481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat 541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa 601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca 661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca 721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat 781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg 841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac 901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac 961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg 1021 cgatcgccat gatggtgcac tgtgctggct gtgagcggcc catcctcgac cgctttctgc 1081 tgaacgtact agaccgcgcg tggcatatca aatgtgttca atgctgcgag tgcaaaacca 1141 acctctcgga gaagtgcttc tcacgggaag gcaagctata ctgtaaaaac gactttttca 1201 ggcgctttgg cacaaagtgc gccggctgcg cgcaaggtat ctctccgagc gacctggtac 1261 ggaaggcccg gagcaaagtc ttccacctca actgcttcac ctgtatggtg tgcaataagc 1321 agctatccac cggagaggag ctctacgtga tcgacgagaa caagtttgtg tgcaaggacg 1381 actacttaag ctcctctagc ctcaaggaag gaagtctcaa ctcggtgtcg tcctgtacgg 1441 accgcagttt gtccccggac ctccaggatc cgttacagga cgaccccaaa gagaccgaca 1501 attcgacctc atcggacaag gaaaccgcta acaacgagaa tgaggaacag aactccggca 1561 ccaaacggcg cggcccgcgc accaccatca aggccaagca gctggagacg ctcaaggcag 1621 ccttcgcagc cacgcccaag cccacgcgcc acattcgcga acagctggca caggagacgg 1681 gcctcaacat gagggtcatt caggtgtggt ttcagaaccg aaggtccaaa gaacgccgca 1741 tgaaacagct gagcgctctg ggcgcgcgga gacacgcctt cttccggagt ccgcggcgca 1801 tgcgtcccct gggcggccgc ttggacgagt ctgagatgtt ggggtctacc ccatacactt 1861 attacggaga ctaccaaagt gactactacg ctccgggagg caattacgat ttcttcgcgc 1921 acggtccgcc gtcacaggcg cagtcccctg ccgactccag cttcctggca gcatcgggac 1981 ctggctcgac gccgctgggc gcgctggaac cgccgctggc cgggcctcac ggcgcggaca 2041 accccagatt caccgacatg atctcgcatc cggacacgcc gagccccgag ccgggcctgc 2101 ccggtgcgct gcaccccatg ccgggagagg tgttcagcgg cgggcccagc ccgcccttcc 2161 ccatgagcgg caccagcggc tacagtggac ccctgtcgca ccccaaccct gagctcaacg 2221 aagcggccgt atggacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc 2281 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc 2341 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg 2401 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta 2461 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg 2521 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa 2581 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa 2641 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc 2701 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc 2761 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg 2821 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac 2881 gtccagacac agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg 2941 cttggcactg tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg 3001 cggccagcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 3061 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 3121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 3181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 3241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 3301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 3361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 3421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 3481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 3541 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 3601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 3661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 3721 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 3781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 3841 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg aggctatggc 3901 agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac ttggggggtg 3961 gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg gtggggtatc 4021 gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc caacaccgtg 4081 cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta ttgtctcctt 4141 ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga gatccccgcg 4201 ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa 4261 ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc 4321 gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc 4381 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc 4441 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcgatccgc cacacccagc 4501 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag 4561 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt gagcctggcg 4621 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga 4681 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg 4741 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc 4801 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc 4861 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg 4921 gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc ggacaggtcg 4981 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag 5041 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga 5101 gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga 5161 tcgatctttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc 5221 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg 5281 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat 5341 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga 5401 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg 5461 ggagcctggg gactttccac accctaactg acacacattc cacagctggt tctttccgcc 5521 tcaggactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 5581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 5641 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 5701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 5761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 5821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 5881 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 5941 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 6001 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 6061 atttaacaaa aatttaacgc gaatttt //