LOCUS       V025538                 7868 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025538
VERSION     V025538
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             872..1579
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1616..3499
                     /label="SNX18(NM_052870)"
                     /note="SNX18(NM_052870)"
                     /gene="SNX18"
     CDS             3506..3532
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3536..3562
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3566..3592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3630..3854
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3900..4328
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4342..4671
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4738..5337
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5511..5632
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5681..5697)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5705..5721
                     /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(5729..5759)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5774..5795
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6083..6668)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6839..7699)
                     /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(7700..7804)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagccac catggtgagc aagggcgagg aggataacat
      901 ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca
      961 cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa
     1021 gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt
     1081 catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct
     1141 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt
     1201 gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg
     1261 cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc
     1321 ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct
     1381 gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa
     1441 gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa
     1501 cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg
     1561 catggacgag ctgtacaagg gaggtggagg tagtggagga ggcggatcaa gcttcatggc
     1621 gctgcgcgcc cgggcgctgt acgacttcag gtcggagaac ccaggagaga tctcgctgcg
     1681 agagcacgag gtgctgagcc tgtgcagcga gcaggacatc gagggctggc tcgagggggt
     1741 caacagccgc ggcgaccgcg gcctcttccc ggcctcctat gtgcaggtga tccgcgcccc
     1801 cgagcctggc ccggcgggag acggcggccc gggcgccccg gcccgctacg ccaatgtgcc
     1861 ccccgggggc ttcgagcccc tgcctgtcgc gccccccgcc tccttcaagc cgccgcctga
     1921 cgccttccag gcgctgctgc agccacagca ggcgccgcct ccgagcacct tccagccgcc
     1981 cggcgcgggc ttcccgtacg gcgggggcgc cctgcagccg tcgcctcagc agctctacgg
     2041 cggctaccag gccagccaag gcagcgatga tgactgggac gacgagtggg acgacagctc
     2101 cacggtggcg gacgagccgg gcgctctggg cagcggagca tacccggacc tcgacggctc
     2161 gtcttcggcg ggtgtgggcg cagccggccg ctaccgcctg tccacgcgct ccgacctgtc
     2221 cctgggttcc cgcggcggct cggtcccccc gcagcaccac ccgtcggggc ccaagagctc
     2281 ggccaccgtg agccgcaacc tcaatcgctt ctccaccttc gtcaagtccg gcggggaggc
     2341 cttcgtgctg ggggaggcgt caggcttcgt gaaggacggg gacaagctgt gcgtggtgct
     2401 ggggccctat ggccccgagt ggcaggagaa cccctacccg ttccagtgca ccatcgacga
     2461 ccccaccaag cagaccaagt tcaagggcat gaagagctac atctcctaca agctggtgcc
     2521 cacgcacacg caggtgccgg tgcatcggcg ctacaagcac ttcgactggc tgtacgcgcg
     2581 cctggcggag aagttcccgg tcatctccgt gccccacctg cccgagaagc aggccaccgg
     2641 ccgcttcgag gaggacttca tctctaagcg caggaagggc ctgatctggt ggatgaacca
     2701 catggccagc cacccagtgc tggcgcagtg cgacgtcttc cagcacttcc tgacgtgccc
     2761 cagcagcacc gacgagaaag cctggaagca gggcaagagg aaggccgaga aggacgagat
     2821 ggtgggcgcc aacttcttcc tgacccttag cacgcccccc gccgctgccc ttgacctgca
     2881 ggaggtggag agcaagatcg acggcttcaa gtgcttcacc aagaagatgg acgacagcgc
     2941 gctgcagctc aaccacacgg ccaacgagtt cgcgcgcaag caggtgaccg gcttcaaaaa
     3001 ggagtatcag aaggtgggcc agtccttccg cggcctcagc caggcctttg agctggacca
     3061 gcaggccttc tcggtgggcc tgaaccaggc tatcgccttc accggagatg cctatgacgc
     3121 cattggcgag ctcttcgcgg agcagcccag gcaggacctg gatcccgtca tggacctatt
     3181 agcgctgtat caggggcatc tggctaactt cccggacatc atccacgttc agaaaggtaa
     3241 agcctggccc ttagagcagg tgatatggag tgtattgtgc aggctgaaag gggcgacttt
     3301 gacagcagta ccactgtggg tttcagaatc atattctaca ggtgaggaag cgagcagaga
     3361 cgtggacgcc tgggtctttt ccctagagtg taagttggat tgctcgacag gcagcttcct
     3421 cctcgagtat cttgcattag ggaatgagta ctctttctcg aaggttcaaa gagtaccttt
     3481 gatgacagtg ctatcatttc tcgagtaccc atacgacgta cctgattatg caacttatcc
     3541 ttatgatgta cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg
     3601 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3661 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3721 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3781 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3841 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3901 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3961 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4021 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4081 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4141 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4201 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4261 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4321 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4441 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4501 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4561 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4621 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4681 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4741 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     4801 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     4861 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4921 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4981 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5041 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5101 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5161 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5221 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5281 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgagcg
     5341 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5401 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5461 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5521 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5581 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5641 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5701 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5761 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5821 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6241 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6481 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6601 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6901 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7021 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7261 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7501 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7681 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7741 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7801 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     7861 cctatggt
//
