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&nbsp;(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
//
