LOCUS       V044880                 6879 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044880
VERSION     V044880
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1559
                     /label="Zfand1(NM_025512)"
                     /note="Zfand1(NM_025512)"
                     /gene="Zfand1"
     CDS             1566..1631
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1655..2243
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2309..2357
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2559..2987
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3001..3330
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3397..4191
                     /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    4367..4488
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4537..4553)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4561..4577
                     /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(4585..4615)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4630..4651
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4939..5527)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5698..6558)
                     /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(6559..6663)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatggc ggagttggac attgggcagc
      781 actgccaggt gcagcactgc cggcagcgag attttcttcc atttgtgtgt gatggttgtt
      841 caggaatatt ttgccttgaa cacagaagca aggactctca tggttgttct gaggtgaatg
      901 tagtcaagga gagaccaaag acagatgaac acaaatctta ctcatgctct ttcaagggct
      961 gcactgacgt agagcttgtg gcagtgatct gtccttactg tgagaagaac ttttgcctga
     1021 gacaccgtca tcagtcggat catgactgtg aaaagctgga agtcgcaaag cctcgaatgg
     1081 ctgccacaca gaaacttgtc agagacattg tcgatgccaa gacaggaggg gcagcaagta
     1141 agggacggaa aggcgcaaaa agtagcggaa cagcggcgaa ggtggcgctg atgaagctaa
     1201 agatgcacgc tgatggggat aagtccctgc cgcagacaga aagaacatac ttccaggtct
     1261 acttgccaaa ggggagcaag gaaaaaagca aagccatgtt cttttgcctg cggtggagca
     1321 tcgggaaggt tgtggacttt gcagcttctc tcgctaatct caggaatgaa aacaacaagc
     1381 tgacagcgaa gaagctgagg ctgtgtcatg tcccttcagg agaagccctg ccattggatc
     1441 acacattgga aaggtggatc acgaaggagg agtgcccttt atataatgga ggaaatgtta
     1501 tcttggaata tcttaacgat gaagaacagt ttttaaaaaa tgttgactct tacttggaag
     1561 gatccgacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     1621 acgatgacaa gtagtgaggg cccgatatct cgacaatcaa cctctggatt acaaaatttg
     1681 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     1741 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     1801 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     1861 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     1921 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     1981 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     2041 gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg
     2101 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     2161 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     2221 ctccctttgg gccgcctccc cgcctggaaa cgggggaggc taactgaaac acggaaggag
     2281 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg
     2341 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     2401 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     2461 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     2521 agcagatctg cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2581 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2641 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2701 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2761 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2821 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2881 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     2941 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3001 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3061 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3121 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3181 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3241 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     3301 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     3361 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3421 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3481 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3541 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     3601 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3661 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3721 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3781 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3841 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3901 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     3961 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4021 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4081 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4141 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4201 tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4261 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4321 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4381 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4441 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4501 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6661 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6721 ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6781 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6841 caaggcttga ccgacaattg catgaagaat ctgcttagg
//
