LOCUS       V040270                 7654 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040270
VERSION     V040270
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             964..3144
                     /label="PLOD1(NM_000302)"
                     /note="PLOD1(NM_000302)"
                     /gene="PLOD1"
     CDS             3151..3216
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3254..3478
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3524..3952
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3966..4295
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4362..5156
                     /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    5330..5451
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5500..5516)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5524..5540
                     /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(5548..5578)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5593..5614
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5902..6487)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6658..7518)
                     /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(7519..7623)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgcggc ccctgctgct actggccctg ctgggctggc tgctgctggc cgaagcgaag
     1021 ggcgacgcca agccggagga caacctttta gtcctcacgg tggccactaa ggagaccgag
     1081 ggattccgtc gcttcaagcg ctcagctcag ttcttcaact acaagatcca ggcgcttggc
     1141 ctaggggagg actggaatgt ggagaagggg acgtcggcag gtggagggca gaaggtccgg
     1201 ctgctgaaga aagctctgga gaagcacgca gacaaggagg atctggtcat tctcttcgca
     1261 gacagctatg acgtgctgtt tgcatcgggg ccccgggagc tcctgaagaa gttccggcag
     1321 gccaggagcc aggtggtctt ctctgctgag gagctcatct acccagaccg caggctggag
     1381 accaagtatc cggtggtgtc cgatggcaag aggttcctgg gctctggagg cttcatcggt
     1441 tatgccccca acctcagcaa actggtggcc gagtgggagg gccaggacag cgacagcgat
     1501 cagctgtttt acaccaagat cttcttggac ccggagaaga gggagcagat caatatcacc
     1561 ctggaccacc gctgccgtat cttccagaac ctggatggag ccttggatga ggtcgtgctc
     1621 aagtttgaaa tgggccatgt gagagcgagg aacctggcct atgacaccct cccggtcctg
     1681 atccatggca acgggccaac caagctgcag ttgaactacc tgggcaacta catcccgcgc
     1741 ttctggacct tcgaaacagg ctgcaccgtg tgtgacgaag gcttgcgcag cctcaagggc
     1801 attggggatg aagctctgcc cacggtcctg gtcggcgtgt tcatcgaaca gcccacgccg
     1861 tttgtgtccc tgttcttcca gcggctcctg cggctccact acccccagaa acacatgcga
     1921 cttttcatcc acaaccacga gcagcaccac aaggctcagg tggaagagtt cctggcacag
     1981 catggcagcg agtaccagtc tgtgaagctg gtgggccctg aggtgcggat ggcgaatgca
     2041 gatgccagga acatgggcgc agacctgtgc cggcaggacc gcagctgcac ctactacttc
     2101 agcgtggatg ctgacgtggc cctgaccgag cccaacagcc tgcggctgct gatccaacag
     2161 aacaagaacg tcattgcccc gctgatgacc cggcatggga ggctgtggtc gaacttctgg
     2221 ggggctctca gtgcagatgg ctactatgcc cgttccgagg actacgtgga cattgtgcag
     2281 gggcggcgtg ttggtgtctg gaatgtgccc tatatttcaa acatctactt gatcaagggc
     2341 agtgccctgc ggggtgagct gcagtcctca gatctcttcc accacagcaa gctggacccc
     2401 gacatggcct tctgtgccaa catccggcag caggatgtgt tcatgttcct gaccaaccgg
     2461 cacacccttg gccatctgct ctccctagac agctaccgca ccacccacct gcacaacgac
     2521 ctctgggagg tgttcagcaa ccccgaggac tggaaggaga agtacatcca ccagaactac
     2581 accaaagccc tggcagggaa gctggtggag acgccctgcc cggatgtcta ttggttcccc
     2641 atcttcacgg aggtggcctg tgatgagctg gtggaggaga tggagcactt tggccagtgg
     2701 tctctgggca acaacaagga caaccgcatc cagggtggct acgagaacgt gccgactatt
     2761 gacatccaca tgaaccagat cggctttgag cgggagtggc acaaattcct gctggagtac
     2821 attgcgccca tgacggagaa gctctacccc ggctactaca ccagggccca gtttgacctg
     2881 gcctttgtcg tccgctacaa gcctgatgag cagccctcac tgatgccaca ccatgatgcc
     2941 tccaccttca ccatcaacat cgccctgaac cgagtcgggg tggattacga gggcgggggc
     3001 tgtcggttcc tgcgctacaa ctgttccatc cgagccccaa ggaagggctg gaccctcatg
     3061 caccctggac gactcacgca ttaccatgag gggctcccca ccaccagggg cacccgctac
     3121 atcgcagtct ccttcgtcga tcccctcgag gactacaaag accatgacgg tgattataaa
     3181 gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccgt ttaaacccgc
     3241 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3301 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3361 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3421 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3481 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3541 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3601 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3661 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3721 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3781 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3841 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3901 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3961 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4021 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     4081 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4141 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4201 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     4261 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     4321 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     4381 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     4441 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4501 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     4561 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4621 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4681 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4741 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4801 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4861 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4921 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4981 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     5041 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     5101 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     5161 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5221 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5281 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5341 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5401 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5461 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5521 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5581 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5641 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5701 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5761 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5821 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5881 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5941 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6001 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6061 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6121 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6181 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6241 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6301 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6361 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6421 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6481 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6541 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6601 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6661 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6721 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6781 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6841 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6901 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6961 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7021 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7081 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7141 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7201 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7261 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7321 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7381 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7441 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7501 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7561 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7621 gcgcacattt ccccgaaaag tgccacctga cgtc
//
