LOCUS       V022428                 9277 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022428
VERSION     V022428
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             932..4756
                     /label="Prdm16(NM_027504)"
                     /note="Prdm16(NM_027504)"
                     /gene="Prdm16"
     CDS             4778..4843
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4872..5096
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      5142..5570
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5584..5913
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5980..6774
                     /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    6950..7071
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7120..7136)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7144..7160
                     /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(7168..7198)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7213..7234
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7522..8110)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8281..9141)
                     /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(9142..9246)
                     /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 gctggctagt
      901 taagcttggt accgagctcg gatccgccac catgcgatcc aaggcgaggg cgaggaagct
      961 agccaaaagt gacggtgacg ttgtaaataa tatgtatgaa cctgacccgg acctgctggc
     1021 cggccagagt gccgaggagg agaccgaaga cggcatcctg tcccccatcc ccatggggcc
     1081 accgtccccc ttccccacca gcgaggactt cactcccaag gagggctcgc cctatgaggc
     1141 tcctgtctac attcctgaag acattccaat cccaccagac ttcgagctac gagagtcctc
     1201 cataccagga gctggcctgg ggatctgggc caagcggaag atggaaatcg gggagaggtt
     1261 tggcccctac gtggtgacgc cccgggccgc actgaaggag gccgactttg gatgggagat
     1321 gctgacggat acagaggtgt catcccagga gagctgcatc aaaaagcaga tctctgaaga
     1381 cttgggtagc gagaagttct gcgtggatgc caatcaggcg gggtctggca gctggctcaa
     1441 gtacatccgt gtagcgtgtt cctgtgatga ccaaaacctc gccatgtgtc agatcaacga
     1501 acagatttac tataaagtca ttaaggacat cgagcctgga gaggaactgt tggtgcatgt
     1561 gaaagaaggt gcctactcct tgggtgtcat ggcccccagc ttggatgagg accccacatt
     1621 ccgctgtgat gagtgtgatg agctcttcca gtgcaggctg gacctgaggc gccacaagaa
     1681 gtacgcgtgc agctctgcag gagcccagct ctacgagggc ctaggggagg aactcaagcc
     1741 cgagggcctt ggcgtgggca gcgacgggca agcgcatgag tgcaaggatt gcgagcggat
     1801 gttccccaac aagtacagct tggagcaaca catgatcgtc cacacggaag agcgtgagta
     1861 caaatgtgac cagtgtccca aggccttcaa ctggaagtcc aacctcatcc gccaccagat
     1921 gtctcacgac agtggcaagc gcttcgaatg tgaaaactgt gtcaaggtgt tcacggaccc
     1981 cagcaacctc cagcgtcaca tccgctcaca gcatgtcggt gcccgggccc atgcctgccc
     2041 tgactgtggc aagaccttcg ccacatcctc tggcctcaaa cagcacaagc atatccacag
     2101 cacggtgaag ccattcatat gcgaggtctg ccacaagtcc tacacgcagt tctccaacct
     2161 gtgccggcac aagcggatgc acgccgactg caggacgcag atcaagtgca aggactgtgg
     2221 gcagatgttc agcactacct cctccctcaa caagcatcgg agattctgcg agggcaagaa
     2281 ccattacacg cctggcagca tcttcacccc aggcctgccc ttgaccccca gccccatgat
     2341 ggacaagaca aaaccctccc cgaccctcaa ccacgggggc ctaggcttca gcgagtactt
     2401 cccctccaga cctcatcctg ggagcctgcc cttctcggct gctcctccgg ccttccccgc
     2461 actcactccg ggcttcccgg gcatctttcc tccatccctg tacccacgac cacctctgct
     2521 acctcccacg ccgctgctca agagccccct gaaccacgcg caggacgcca agctacccag
     2581 cccgctggga aacccagccc tgccccttgt ctccgcggtc agcaatagca gccagggtgc
     2641 cacagcggcc accgggtcag aggagaaatt tgatggccgc ttggaagacg catatgcgga
     2701 gaaggtcaaa aataggagcc ctgacatgtc ggatggcagt gactttgagg atatcaacac
     2761 cacgaccggg acagacttgg acactaccac gggcacgggg tcagacctgg acagcgacct
     2821 ggacagtgac agagacaaag gcaaggacaa ggggaagcca gtggagagca aacctgagtt
     2881 tgggggtgca tctgtgcccc ctggggccat gaacagtgtg gccgaggtac cggccttcta
     2941 ctcacagcat tccttcttcc cgccacccga ggaacagctg ctgacggcct cgggagctgc
     3001 cggcgactcc atcaaggcca tcgcgtccat cgcggagaaa tacttcggtc ctggcttcat
     3061 gagcatgcag gagaagaagc tgggctcact accctaccac tccgtgttcc ccttccagtt
     3121 cctgcctaac tttccccact ccctctaccc ctttacggac cgagccctcg cccacaactt
     3181 gctggtcaag gctgagccaa agtcaccccg ggatgccctc aaggtgggcg gccccagtgc
     3241 ggagtgcccc ttcgacctca ccaccaaacc aaaagaggcc aaacccgccc tgctcgcacc
     3301 caaggtcccc ctcatcccct catctggcga ggaacagcca ctggacctga gcatcggcag
     3361 cagggccagg gcaagccaga acggaggtgg ccgtgagccg cggaagaacc acgtctacgg
     3421 tgaacggaag ccgggggtca gcgaggggct gcctaaggtg tgcccagcac agctgcccca
     3481 gcagccctcc ttgcattatg ctaagccttc accgttcttc atggatccca tctacagggt
     3541 agaaaagcgg aaggtggcag accctgtggg agtcctgaaa gagaagtacc tgcggccgtc
     3601 cccacttctg ttccaccccc agatgtcagc catagaaacc atgacggaga agctggagag
     3661 ctttgcagcc atgaaggccg actcaggcag ctccctgcag cccctgcctc accacccgtt
     3721 caacttccgc tccccacccc caacgctctc ggatcccatc ctcaggaagg ggaaggagag
     3781 atacacgtgc aggtactgtg gcaagatctt ccccagatct gcaaatctca caagacatct
     3841 gaggacacac acaggggagc agccatacag gtgcaagtac tgtgaccggt cattcagcat
     3901 ctcctccaac ctccagcggc acgtgaggaa catccacaac aaagagaagc cgttcaagtg
     3961 ccatctgtgc aaccgctgct tcgggcagca gaccaaccta gaccggcacc tgaagaagca
     4021 cgaacacgag ggcgcaccag tgagccagca ctccggggtg ctcacgaacc acctgggcac
     4081 cagcgcctcc tcccccacct ccgagtcgga caaccatgca cttttagatg agaaggaaga
     4141 ttcttacttc tccgagatcc gaaacttcat cgccaacagc gagatgaacc aggcatccac
     4201 tcgaatggac aaacggcctg agatccaaga cctggacagc aacccaccgt gtccaggctc
     4261 agccagtgca aagccagagg acgtagagga ggaggaagag gaggagctgg aggaagagga
     4321 tgatgacagc ttagccggga agtcacagga ggacacggtg tcccccacac ctgagcccca
     4381 aggagtctat gaagatgaag aggatgagga accacccagc ctgaccatgg gctttgacca
     4441 tacccggagg tgtgttgagg agcgaggagg cggcctgtta gctttggagc cgacgccgac
     4501 ctttgggaag gggctggatc tccgcagagc agctgaggaa gcatttgaag ttaaagatgt
     4561 gcttaattcc accttagatt ctgaggtttt aaaacaaacc ctgtacaggc aggctaagaa
     4621 ccaggcatat gcaatgatgc tgtccctctc tgaagacact cctctccacg ccccctccca
     4681 gagctcactg gatgcttggt tgaacatcac aggaccctcg tcagagtccg gagcctttaa
     4741 ccccatcaac cacctcctcg agtctagagg gcccttcgac tacaaagacc atgacggtga
     4801 ttataaagat catgacatcg actacaagga tgacgatgac aagtgagttt aaacccgctg
     4861 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     4921 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     4981 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     5041 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     5101 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     5161 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5221 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5281 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5341 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5401 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5461 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5521 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     5581 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     5641 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     5701 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     5821 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     5881 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     5941 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6001 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6061 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6121 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6181 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6241 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6301 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6361 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6421 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6481 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     6541 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6601 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6661 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6721 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6781 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     6841 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     6901 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     6961 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     7021 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     7081 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     7141 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7201 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7261 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7321 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7381 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7441 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7501 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7561 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7621 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7681 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7741 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7801 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7861 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7921 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     7981 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     8041 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     8101 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     8161 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     8221 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8281 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8341 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8401 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8461 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8521 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8581 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8641 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8701 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8761 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8821 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8881 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8941 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     9001 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     9061 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     9121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     9181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9241 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
