LOCUS       V037612                 6767 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037612
VERSION     V037612
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             955..1926
                     /label="TMEM68(NM_001286657)"
                     /note="TMEM68(NM_001286657)"
                     /gene="TMEM68"
     CDS             1963..2679
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             2689..2718
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2734..2751
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2780..3004
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3050..3478
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3492..3822
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        3870..3917
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             3936..4310
                     /codon_start=1
                     /gene="Sh&nbsp;ble from&nbsp;Streptoalloteichus
                     hindustanus"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    4440..4561
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4610..4626)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4634..4650
                     /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(4658..4688)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4703..4724
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5012..5600)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5771..6631)
                     /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(6632..6736)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gatccactag tccagtgtgg tggaattcgc caccatgata
      961 gacaaaaatc aaacctgtgg tgtaggacag gattctgtgc cctatatgat ttgtctgatt
     1021 cacatactcg aagaatggtt tggtgtggag cagttggagg actatttgaa ttttgcaaac
     1081 tatctcttgt gggtttttac accactaata cttttaatac ttccttactt tactatcttt
     1141 cttctctacc ttactattat tttcttacac atttataaga gaaagaatgt attgaaagaa
     1201 gcctactctc ataatttatg ggatggtgca aggaaaacag tggcaactct gtgggatgga
     1261 catgcagccg tttggcatgg ttatgaagtt catggaatgg aaaaaatacc agaagatgga
     1321 ccagcactta taatttttta tcatggagct attcctatag atttttacta tttcatggct
     1381 aaaatattta tacacaaagg cagaacttgc cgagtagtag ctgatcactt tgtctttaaa
     1441 attccagggt ttagtttatt actggatgtg ttttgtgctc tacatggacc aagagaaaaa
     1501 tgtgttgaaa ttctgaggag tggccacttg ttagctatct caccaggtgg agttcgagaa
     1561 gccctaatta gtgatgaaac ttataacatc gtatggggtc atcgcagagg ctttgctcag
     1621 gttgcaattg atgcaaaagt gcccattatt cctatgttta cacaaaatat tcgagaagga
     1681 tttagatcac ttggaggaac aaggttattt aggtggcttt atgaaaaatt ccgctatcca
     1741 tttgctccaa tgtatggagg ttttccagtg aagttacgga cctatttagg cgaccccatt
     1801 ccgtatgacc cacagataac agcggaagaa ttagctgaaa agacgaagaa tgctgttcaa
     1861 gctttgattg ataagcacca aagaatacca ggaaacatta tgagtgcttt gttagaacgt
     1921 tttcatggat ccggcggtgg aggaagcggc ggaggtggat ctatggtgag caagggcgag
     1981 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2041 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2101 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     2161 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     2221 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2281 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2341 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2401 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     2461 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     2521 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     2581 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2641 gccgccggga tcactctcgg catggacgag ctgtacaagc cgcggttcga acaaaaactc
     2701 atctcagaag aggatctgaa tatgcatacc ggtcatcatc accatcacca ttgagtttaa
     2761 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     2821 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2881 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2941 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3001 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3061 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3121 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3181 tttccccgtc aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg
     3241 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3301 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3361 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3421 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3481 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3841 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3901 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     3961 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     4021 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4081 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4141 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4201 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4261 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
