LOCUS       V043330                 7119 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043330
VERSION     V043330
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..1845
                     /label="Fos(NM_010234)"
                     /note="Fos(NM_010234)"
                     /gene="Fos"
     CDS             1852..1875
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    1895..2483
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2549..2597
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2799..3227
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3241..3570
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3637..4431
                     /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    4607..4728
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4777..4793)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4801..4817
                     /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(4825..4855)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4870..4891
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5179..5767)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5938..6798)
                     /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(6799..6903)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgat gttctcgggt
      721 ttcaacgccg actacgaggc gtcatcctcc cgctgcagta gcgcctcccc ggccggggac
      781 agcctttcct actaccattc cccagccgac tccttctcca gcatgggctc tcctgtcaac
      841 acacaggact tttgcgcaga tctgtccgtc tctagtgcca actttatccc cacggtgaca
      901 gccatctcca ccagcccaga cctgcagtgg ctggtgcagc ccactctggt ctcctccgtg
      961 gccccatcgc agaccagagc gccccatcct tacggactcc ccacccagtc tgctggggct
     1021 tacgccagag cgggaatggt gaagaccgtg tcaggaggca gagcgcagag catcggcaga
     1081 aggggcaaag tagagcagct atctcctgaa gaggaagaga aacggagaat ccgaagggaa
     1141 cggaataaga tggctgcagc caagtgccgg aatcggagga gggagctgac agatacactc
     1201 caagcggaga cagatcaact tgaagatgag aagtctgcgt tgcagactga gattgccaat
     1261 ctgctgaaag agaaggaaaa actggagttt attttggcag cccaccgacc tgcctgcaag
     1321 atccccgatg accttggctt cccagaggag atgtctgtgg cctccctgga tttgactgga
     1381 ggtctgcctg aggcttccac cccagagtct gaggaggcct tcaccctgcc ccttctcaac
     1441 gaccctgagc ccaagccatc cttggagcca gtcaagagca tcagcaacgt ggagctgaag
     1501 gcagaaccct ttgatgactt cttgtttccg gcatcatcta ggcccagtgg ctcagagacc
     1561 tcccgctctg tgccagatgt ggacctgtcc ggttccttct atgcagcaga ctgggagcct
     1621 ctgcacagca attccttggg gatggggccc atggtcacag agctggagcc cctgtgtact
     1681 cccgtggtca cctgtactcc gggctgcact acttacacgt cttcctttgt cttcacctac
     1741 cctgaagctg actccttccc aagctgtgcc gctgcccacc gaaagggcag cagcagcaac
     1801 gagccctcct ccgactccct gagctcaccc acgctgctgg ccctgctcga gcaccatcac
     1861 catcaccatc accattaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     1921 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1981 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     2041 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     2101 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     2161 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     2221 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2281 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2341 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2401 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2461 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     2521 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2581 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     2641 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     2701 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     2761 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2821 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2881 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2941 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3001 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3061 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3121 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3181 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3241 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3301 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3361 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3421 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3481 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3541 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3601 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3661 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3721 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3781 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3841 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3901 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3961 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4021 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4081 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4141 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4201 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4261 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4321 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4381 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4441 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4501 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4561 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4621 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4681 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4741 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4801 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4861 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4921 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4981 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5041 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5101 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5161 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5221 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5281 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5341 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5401 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5461 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5521 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5581 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5641 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5701 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5761 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5821 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5881 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5941 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6001 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6061 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6541 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6901 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6961 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     7021 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     7081 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
