LOCUS       V025138                 6957 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025138
VERSION     V025138
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..1637
                     /label="RARRES1(NM_206963)"
                     /note="RARRES1(NM_206963)"
                     /gene="RARRES1"
     CDS             1644..1709
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1733..2321
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2387..2435
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2637..3065
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3079..3408
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3475..4269
                     /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    4445..4566
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4615..4631)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4639..4655
                     /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(4663..4693)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4708..4729
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5017..5605)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5776..6636)
                     /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(6637..6741)
                     /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 ccaccatgca gccccgccgg caacggctgc
      781 ctgctccctg gtccgggccc aggggcccgc gccccaccgc cccgctgctc gcgctgctgc
      841 tgttgctcgc cccggtggcg gcgcccgcgg ggtccgggga ccccgacgac cctgggcagc
      901 ctcaggatgc tggggtcccg cgcaggctcc tgcagcaggc ggcgcgcgcg gcgcttcact
      961 tcttcaactt ccggtccggc tcgcccagcg cgctacgagt gctggccgag gtgcaggagg
     1021 gccgcgcgtg gattaatcca aaagagggat gtaaagttca cgtggtcttc agcacagagc
     1081 gctacaaccc agagtcttta cttcaggaag gtgagggacg tttggggaaa tgttctgctc
     1141 gagtgttttt caagaatcag aaacccagac caaccatcaa tgtaacttgt acacggctca
     1201 tcgagaaaaa gaaaagacaa caagaggatt acctgcttta caagcaaatg aagcaactga
     1261 aaaacccctt ggaaatagtc agcatacctg ataatcatgg acatattgat ccctctctga
     1321 gactcatctg ggatttggct ttccttggaa gctcttacgt gatgtgggaa atgacaacac
     1381 aggtgtcaca ctactacttg gcacagctca ctagtgtgag gcagtggaaa actaatgatg
     1441 atacaattga ttttgattat actgttctac ttcatgaatt atcaacacag gaaataattc
     1501 cctgtcgcat tcacttggtc tggtaccctg gcaaacctct taaagtgaag taccactgtc
     1561 aagagctaca gacaccagaa gaagcctccg gaactgaaga aggatcagct gtagtaccaa
     1621 cagagcttag taatttcgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1681 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1741 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1801 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1861 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1921 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1981 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2041 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2101 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2161 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2221 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2281 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2341 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2401 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2461 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2521 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2581 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2641 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2701 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2761 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2821 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2881 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2941 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3001 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3061 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3121 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3181 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3241 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3301 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3361 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3421 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3481 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3541 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3601 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3661 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3721 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3781 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3841 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3901 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3961 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4021 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4081 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4141 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4201 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4261 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4321 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4381 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4441 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4561 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4621 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4681 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4741 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4801 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4861 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4921 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4981 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5041 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5101 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5161 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5221 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5281 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5341 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5401 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5461 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5521 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5581 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5641 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5701 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5761 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5821 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5881 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5941 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6001 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6061 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6121 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6181 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6241 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6301 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6361 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6421 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6481 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6541 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6601 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6661 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6721 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6781 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6841 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6901 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
