LOCUS       V024435                 8327 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024435
VERSION     V024435
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             874..1590
                     /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             1660..3699
                     /label="COL10A1(NM_001424106)"
                     /note="COL10A1(NM_001424106)"
                     /gene="COL10A1"
     CDS             3706..3732
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3736..3762
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3766..3792
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3894..4118
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4164..4592
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4606..4935
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5002..5796
                     /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    5970..6091
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6140..6156)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6164..6180
                     /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(6188..6218)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6233..6254
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6542..7127)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7298..8158)
                     /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(8159..8263)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgcc accatggtga gcaagggcga ggagctgttc
      901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1561 atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagg cggcggaggg
     1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tgctgccaca aatacccttt
     1681 ttgctgctag tatccttgaa cttggttcat ggagtgtttt acgctgaacg ataccaaatg
     1741 cccacaggca taaaaggccc actacccaac accaagacac agttcttcat tccctacacc
     1801 ataaagagta aaggtatagc agtaagagga gagcaaggta ctcctggtcc accaggccct
     1861 gctggacctc gagggcaccc aggtccttct ggaccaccag gaaaaccagg ctacggaagt
     1921 cctggactcc aaggagagcc agggttgcca ggaccaccgg gaccatcagc tgtagggaaa
     1981 ccaggtgtgc caggactccc aggaaaacca ggagagagag gaccatatgg accaaaagga
     2041 gatgttggac cagctggcct accaggaccc cggggcccac caggaccacc tggaatccct
     2101 ggaccggctg gaatttctgt gccaggaaaa cctggacaac agggacccac aggagcccca
     2161 ggacccaggg gctttcctgg agaaaagggt gcaccaggag tccctggtat gaatggacag
     2221 aaaggggaaa tgggatatgg tgctcctggt cgtccaggtg agaggggtct tccaggccct
     2281 cagggtccca caggaccatc tggccctcct ggagtgggaa aaagaggtga aaatggggtt
     2341 ccaggacagc caggcatcaa aggtgataga ggttttccgg gagaaatggg accaattggc
     2401 ccaccaggtc cccaaggccc tcctggggaa cgagggccag aaggcattgg aaagccagga
     2461 gctgctggag ccccaggcca gccagggatt ccaggaacaa aaggtctccc tggggctcca
     2521 ggaatagctg ggcccccagg gcctcctggc tttgggaaac caggcttgcc aggcctgaag
     2581 ggagaaagag gacctgctgg ccttcctggg ggtccaggtg ccaaagggga acaagggcca
     2641 gcaggtcttc ctgggaagcc aggtctgact ggaccccctg ggaatatggg accccaagga
     2701 ccaaaaggca tcccgggtag ccatggtctc ccaggcccta aaggtgagac agggccagct
     2761 gggcctgcag gataccctgg ggctaagggt gaaaggggtt cccctgggtc agatggaaaa
     2821 ccagggtacc caggaaaacc aggtctcgat ggtcctaagg gtaacccagg gttaccaggt
     2881 ccaaaaggtg atcctggagt tggaggacct cctggtctcc caggccctgt gggcccagca
     2941 ggagcaaagg gaatgcccgg acacaatgga gaggctggcc caagaggtgc ccctggaata
     3001 ccaggtacta gaggccctat tgggccacca ggcattccag gattccctgg gtctaaaggg
     3061 gatccaggaa gtcccggtcc tcctggccca gctggcatag caactaaggg cctcaatgga
     3121 cccaccgggc caccagggcc tccaggtcca agaggccact ctggagagcc tggtcttcca
     3181 gggccccctg ggcctccagg cccaccaggt caagcagtca tgcctgaggg ttttataaag
     3241 gcaggccaaa ggcccagtct ttctgggacc cctcttgtta gtgccaacca gggggtaaca
     3301 ggaatgcctg tgtctgcttt tactgttatt ctctccaaag cttacccagc aataggaact
     3361 cccataccat ttgataaaat tttgtataac aggcaacagc attatgaccc aaggactgga
     3421 atctttactt gtcagatacc aggaatatac tatttttcat accacgtgca tgtgaaaggg
     3481 actcatgttt gggtaggcct gtataagaat ggcacccctg taatgtacac ctatgatgaa
     3541 tacaccaaag gctacctgga tcaggcttca gggagtgcca tcatcgatct cacagaaaat
     3601 gaccaggtgt ggctccagct tcccaatgcc gagtcaaatg gcctatactc ctctgagtat
     3661 gtccactcct ctttctcagg attcctagtg gctccaatgg gtacctaccc atacgacgta
     3721 cctgattatg caacttatcc ttatgatgta cctgattatg ctagctatcc ttatgatgtc
     3781 ccagactacg cataggatcc actagtccag tgtggtggaa ttctgcagat atccagcaca
     3841 gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3901 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3961 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4021 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4081 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4141 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4201 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     4261 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4321 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     4381 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4441 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4501 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4561 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     4621 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4681 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4741 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4801 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4861 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4921 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4981 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     5041 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     5101 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     5161 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     5221 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     5281 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     5341 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     5401 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     5461 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     5521 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     5581 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     5641 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5701 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5761 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5821 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5881 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5941 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     6001 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     6061 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     6121 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     6181 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     6241 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     6301 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6361 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6421 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6481 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6541 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6601 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6661 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6721 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6781 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6841 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6901 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6961 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7021 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt
     7081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8281 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
