LOCUS       V028223                 6411 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028223
VERSION     V028223
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(941..1801)
                     /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(1802..1906)
                     /gene="bla"
                     /label="AmpR promoter"
     enhancer        2172..2551
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2552..2755
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     protein_bind    2757..2775
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    2778..2796
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     CDS             2963..4282
                     /label="SQSTM1(NM_003900)"
                     /note="SQSTM1(NM_003900)"
                     /gene="SQSTM1"
     CDS             4292..4315
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    4362..4586
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4632..5060
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5074..5403
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     promoter        5451..5498
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             5517..5891
                     /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    6021..6142
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6191..6207)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6215..6231
                     /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(6239..6269)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6284..6305
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctccc
     2761 tatcagtgat agagatctcc ctatcagtga tagagatcgt cgacgagctc gtttagtgaa
     2821 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     2881 ccgatccagc ctccggactc tagcgtttaa acttaagctt ggtaccgagc tcggatccac
     2941 tagtccagtg tggtggaatt ctatggcgtc gctcaccgtg aaggcctacc ttctgggcaa
     3001 ggaggacgcg gcgcgcgaga ttcgccgctt cagcttctgc tgcagccccg agcctgaggc
     3061 ggaagccgag gctgcggcgg gtccgggacc ctgcgagcgg ctgctgagcc gggtggccgc
     3121 cctgttcccc gcgctgcggc ctggcggctt ccaggcgcac taccgcgatg aggacgggga
     3181 cttggttgcc ttttccagtg acgaggaatt gacaatggcc atgtcctacg tgaaggatga
     3241 catcttccga atctacatta aagagaaaaa agagtgccgg cgggaccacc gcccaccgtg
     3301 tgctcaggag gcgccccgca acatggtgca ccccaatgtg atctgcgatg gctgcaatgg
     3361 gcctgtggta ggaacccgct acaagtgcag cgtctgccca gactacgact tgtgtagcgt
     3421 ctgcgaggga aagggcttgc accgggggca caccaagctc gcattcccca gccccttcgg
     3481 gcacctgtct gagggcttct cgcacagccg ctggctccgg aaggtgaaac acggacactt
     3541 cgggtggcca ggatgggaaa tgggtccacc aggaaactgg agcccacgtc ctcctcgtgc
     3601 aggggaggcc cgccctggcc ccacggcaga atcagcttct ggtccatcgg aggatccgag
     3661 tgtgaatttc ctgaagaacg ttggggagag tgtggcagct gcccttagcc ctctgggcat
     3721 tgaagttgat atcgatgtgg agcacggagg gaaaagaagc cgcctgaccc ccgtctctcc
     3781 agagagttcc agcacagagg agaagagcag ctcacagcca agcagctgct gctctgatcc
     3841 cagcaagccg ggtgggaatg ttgagggcgc cacgcagtct ctggcggagc agatgagaaa
     3901 gatcgccttg gagtccgagg ggcgccctga ggaacagatg gagtcggata actgttcagg
     3961 aggagatgat gactggaccc atctgtcttc aaaagaagtg gacccgtcta caggtgaact
     4021 ccagtcccta cagatgccag aatccgaagg gccaagctct ctggacccct cccaggaggg
     4081 acccacaggg ctgaaggaag ctgccttgta cccacatctc ccgccagagg ctgacccgcg
     4141 gctgattgag tccctctccc agatgctgtc catgggcttc tctgatgaag gcggctggct
     4201 caccaggctc ctgcagacca agaactatga catcggagcg gctctggaca ccatccagta
     4261 ttcaaagcat cccccgccgt tgcgcggccg cgactacaag gatgacgatg acaagtgact
     4321 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4381 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4441 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4501 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4561 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4621 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4681 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4741 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4801 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4861 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4921 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4981 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5041 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5101 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5221 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5281 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5341 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5401 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt
     5461 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg
     5521 ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt
     5581 tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc gccggtgtgg
     5641 tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtggtg ccggacaaca
     5701 ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg
     5761 tgtccacgaa cttccgggac gcctccgggc cggccatgac cgagatcggc gagcagccgt
     5821 gggggcggga gttcgccctg cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg
     5881 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5941 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6001 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     6061 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6121 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     6181 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6241 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6301 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6361 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc t
//
