LOCUS       V045719                 9939 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045719
VERSION     V045719
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..4583
                     /label="PRDM16(NM_022114)"
                     /note="PRDM16(NM_022114)"
                     /gene="PRDM16"
     CDS             4626..4691
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4715..5303
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    5369..5417
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5619..6047
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        6061..6390
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             6457..7251
                     /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    7427..7548
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7597..7613)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7621..7637
                     /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(7645..7675)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7690..7711
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7999..8587)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8758..9618)
                     /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(9619..9723)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcg atccaaggcg agggcgagga
      781 agctagccaa aagtgacggt gacgttgtaa ataatatgta tgagcccaac cgggacctgc
      841 tggccagcca cagcgcggag gacgaggccg aggacagtgc catgtcgccc atccccgtgg
      901 ggccaccgtc ccccttcccc accagcgagg acttcacccc caaggagggc tcgccgtacg
      961 aggcccctgt ctacattcct gaagacattc cgatcccagc agacttcgag ctccgagagt
     1021 cctccatccc aggggctggc ctgggggtct gggccaagag gaagatggaa gccggggaga
     1081 ggctgggccc ctgcgtggtg gtgccccggg cggcggcaaa ggagacagac ttcggatggg
     1141 agcaaatact gacggacgtg gaagtgtcgc cccaggaagg ctgcatcaca aagatctccg
     1201 aagacctggg cagtgagaag ttctgcgtgg atgcaaatca ggcgggggct ggcagctggc
     1261 tcaagtacat ccgtgtggcg tgctcctgcg atgaccagaa cctcaccatg tgtcagatca
     1321 gtgagcagat ttactataaa gtcattaagg acattgagcc aggtgaggag ctgctggtgc
     1381 acgtgaagga aggcgtctac cccctgggca cagtgccgcc cggcctggac gaggagccca
     1441 cgttccgctg tgacgagtgt gacgaactct tccagtccaa gctggacctg cggcgccata
     1501 agaagtacac gtgtggctca gtgggggctg cgctctacga gggcctggct gaggagctca
     1561 agcccgaggg ccttggcggt ggcagcggcc aagcccacga gtgcaaggac tgcgagcgga
     1621 tgttccccaa caagtacagc ctggagcagc acatggtcat ccacacggag gagcgcgagt
     1681 acaaatgcga ccagtgtccc aaggccttca actggaagtc caacctcatc cgccaccaga
     1741 tgtcccacga cagcggcaaa cgcttcgaat gtgaaaactg cgtgaaggtg ttcacggacc
     1801 ccagcaacct tcagcggcac atccgctcgc agcacgtggg cgctcgggcc cacgcctgcc
     1861 ccgactgcgg gaagaccttc gccacgtcct ccggcctcaa gcagcacaag catatccaca
     1921 gcacggtgaa gcctttcata tgtgaggtct gccacaagtc ctacacgcag ttctccaacc
     1981 tgtgccggca caagcggatg cacgccgact gccgcacgca gatcaagtgc aaggactgtg
     2041 gccagatgtt cagcactacc tcctccctca acaagcaccg gcgcttctgc gagggcaaga
     2101 accattacac gccgggcggc atctttgccc cgggcctgcc cttgaccccc agccccatga
     2161 tggacaaggc aaaaccctcc cccagcctca atcacgccag cctgggcttc aacgagtact
     2221 ttccctccag gccgcacccg gggagcctgc ccttctccac ggcgcctccc acgttccccg
     2281 cactcacccc cggcttcccg ggcatcttcc ctccatcctt gtacccccgg ccgcctctgc
     2341 tacctcccac atcgctgctc aagagccccc tgaaccacac ccaggacgcc aagctcccca
     2401 gtcccctggg gaacccagcc ctgcccctgg tctccgccgt cagcaacagc agccagggca
     2461 cgacggcagc tgcggggccc gaggagaagt tcgagagccg cctggaggac tcctgtgtgg
     2521 agaagctgaa gaccaggagc agcgacatgt cggacggcag tgactttgag gacgtcaaca
     2581 ccaccacggg gaccgacctg gacacgacca cggggacggg ctcggacctg gacagcgacg
     2641 tggacagcga ccctgacaag gacaagggca agggcaagtc cgccgagggc cagcccaagt
     2701 ttgggggcgg cttggcgccc ccgggggccc cgaacagcgt ggccgaggtg cctgtcttct
     2761 attcccagca ctcattcttc ccgccacccg acgagcagct gctgactgca acgggcgccg
     2821 ccggggactc catcaaggcc atcgcatcca ttgccgagaa gtactttggc cccggcttca
     2881 tggggatgca ggagaagaag ctgggctcgc tcccctacca ctcggcgttc cccttccagt
     2941 tcctgcccaa cttcccccac tccctttacc ccttcacgga ccgagccctc gcccacaact
     3001 tgctggtcaa ggccgagcca aagtcacccc gggacgccct caaggtgggc ggccccagtg
     3061 ccgagtgccc ctttgatctc accaccaagc ccaaagacgt gaagcccatc ctgcccatgc
     3121 ccaagggccc ctcggccccc gcatccggcg aggagcagcc gctggacctg agcatcggca
     3181 gccgggcccg tgccagccaa aacggcggcg ggcgggagcc ccgcaagaac cacgtctatg
     3241 gggaacgcaa gctgggcgcc ggcgaggggc tgccccaggt gtgcccggcg cggatgcccc
     3301 agcagccccc gctccactac gccaagccct cgcccttctt catggacccc atctacagca
     3361 gggtagaaaa gcggaaggtc acagaccccg tgggagccct gaaggagaag tacctgcggc
     3421 cgtccccgct gctcttccac ccccagatgt cagccataga gaccatgaca gagaagctgg
     3481 agagctttgc agccatgaag gcggactcgg gcagctccct gcagcccctc ccccaccacc
     3541 ccttcaactt ccggtcccca cccccaacgc tctccgaccc catcctcagg aagggcaagg
     3601 agcgatacac gtgcaggtac tgtgggaaga tcttccccag atcagccaat ctcaccagac
     3661 acctgaggac gcacactggg gagcagccgt acaggtgtaa gtactgcgac cgctccttca
     3721 gcatctcttc gaacctccag cggcacgtcc ggaacatcca caacaaggag aagcctttca
     3781 agtgccacct gtgcaaccgc tgcttcgggc agcagaccaa cctggaccgg cacctcaaga
     3841 agcacgagca cgagaacgca ccagtgagcc agcaccccgg ggtcctcacg aaccacctgg
     3901 ggaccagcgc gtcctctccc acctcagagt cggacaacca cgcactttta gacgagaaag
     3961 aagactctta tttctcggaa atcagaaact ttattgccaa tagtgagatg aaccaagcat
     4021 caacgcgaac agagaaacgg gcggacatgc agatcgtgga cggcagtgcc cagtgtccag
     4081 gcctagccag tgagaagcag gaggacgtgg aggaggagga cgacgatgac ctggaggagg
     4141 acgatgagga cagcctggcc gggaagtcgc aggatgacac cgtgtccccc gcacccgagc
     4201 cccaggccgc ctacgaggat gaggaggatg aggagccagc cgcctccctg gccgtgggct
     4261 ttgaccacac ccgaaggtgt gctgaggacc acgaaggcgg tctgttagct ttggagccga
     4321 tgccgacttt tgggaagggg ctggacctcc gcagagcagc tgaggaagca tttgaagtta
     4381 aagatgtgct taattccacc ttagattctg aggctttaaa acatacactg tgcaggcagg
     4441 ctaagaacca ggcatatgca atgatgctgt ccctttccga agacactcct ctccacaccc
     4501 cctcccaggg ttctctggac gcttggttga aggtcactgg agccacgtcg gagtctggag
     4561 catttcaccc catcaaccac ctctgtatat atgtatatat gtatatatgt atatatgtat
     4621 atagcgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     4681 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     4741 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4801 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4861 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4921 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4981 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     5041 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     5101 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     5161 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     5221 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     5281 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     5341 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     5401 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5461 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5521 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5581 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5641 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5701 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5761 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5821 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5881 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5941 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     6001 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     6061 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     6121 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     6181 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     6241 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     6301 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     6361 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     6421 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     6481 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     6541 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     6601 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     6661 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     6721 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     6781 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     6841 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     6901 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     6961 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     7021 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     7081 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     7141 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     7201 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     7261 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     7321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     7381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     7441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     7501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     7561 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8161 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     8221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8401 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     8461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8581 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8641 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8701 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8761 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8821 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8881 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8941 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     9001 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     9061 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     9121 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     9181 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9241 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9301 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9361 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9421 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9481 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9541 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9601 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9661 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9721 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9781 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9841 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9901 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
