LOCUS       V041122                 8177 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V041122
VERSION     V041122
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             908..3121
                     /label="Il12rb1(NM_008353)"
                     /note="Il12rb1(NM_008353)"
                     /gene="Il12rb1"
     CDS             3128..3184
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             3185..3895
                     /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"
     polyA_signal    3939..4163
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4209..4637
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4651..4980
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5047..5646
                     /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    5820..5941
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5990..6006)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6014..6030
                     /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(6038..6068)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6083..6104
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6392..6977)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7148..8008)
                     /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(8009..8113)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gacatgatgg ggctggcagg gaccagcaaa cacatcacct tcctcctgct
      961 ttgccagcta ggcgcctcag gcccaggtga tggctgctgc gttgagaaga catcgttccc
     1021 agagggagcc tcaggctcac ccttaggacc caggaacttg agttgctaca gggtttccaa
     1081 gacagactat gagtgctcct ggcagtatga tggccctgag gacaatgttt ctcacgtcct
     1141 gtggtgctgc tttgtccctc cgaaccatac ccacaccggc caggagcgct gccgctactt
     1201 ctcctcaggc ccagaccgca ctgtgcagtt ctgggaacag gacggtatcc ctgtgctgtc
     1261 caaggtcaac ttctgggtgg agtctcggct tgggaaccga accatgaagt cccagaagat
     1321 atcccagtac ctgtacaact ggaccaagac gacccctccc ctgggacaca tcaaggtgtc
     1381 acaatcacac cggcagttgc gaatggactg gaatgtgtct gaagaggccg gtgctgaggt
     1441 acagttcagg cgccgtatgc ccacaacgaa ttggaccttg ggtgactgcg gacctcaggt
     1501 taactctggc tcaggtgtgc ttggtgacat tcgtgggagc atgtctgagt cctgcctctg
     1561 cccttctgag aacatggccc aagagatcca gatacggagg aggaggcggc tctcctcagg
     1621 agcccctgga ggtccctgga gtgattggag catgcctgtg tgtgttccac ctgaagtcct
     1681 tccccaggcc aagattaagt tcttggtgga gcccctgaac caaggtggaa ggaggcgtct
     1741 aaccatgcaa ggacagtcac cacagctggc agtcccggaa ggctgccgag gcaggcctgg
     1801 tgcgcaggtg aagaagcact tggtgctggt gcgcatgctg tcctgcaggt gccaggctca
     1861 gacctcgaag accgtgcccc tgggcaagaa gctgaacctc tccggggcca cctatgacct
     1921 gaatgtgctc gccaaaactc gtttcggtcg cagcaccatc cagaagtggc accttcctgc
     1981 ccaagagctc acagagacaa gagccctgaa tgtcagcgtg ggaggcaaca tgacatccat
     2041 gcagtgggca gcccaggctc ccggcaccac ctactgcctt gagtggcagc catggttcca
     2101 gcacaggaac cacacacact gtaccctgat tgtaccagaa gaagaggatc ctgccaagat
     2161 ggtgacacac agctggagct ctaaacctac cctggagcag gaggaatgtt accgcatcac
     2221 agtgttcgcc tccaagaacc ccaagaatcc catgctgtgg gccacagtcc tgtccagtta
     2281 ctactttggg ggtaacgcct cgagagccgg tactccacga cacgtgtcgg tgaggaacca
     2341 aaccggggac tcggtgtccg tggagtggac ggcgtcacag ctgagcacct gcccgggggt
     2401 cctgacgcaa tacgtcgtgc gctgcgaggc tgaagacggc gcgtgggagt cagagtggct
     2461 cgtgccaccc actaaaaccc aagtgacact tgacggactg cgcagccgag taatgtacaa
     2521 ggttcaggtg cgagccgaca ctgcgaggct cccgggtgcc tggagtcatc cccagcgctt
     2581 tagctttgag gtgcagattt cccgtttatc catcattttc gcgtctctgg gaagcttcgc
     2641 cagcgtcctc ctcgtgggca gtctcggata cattggctta aacagggccg cctggcactt
     2701 gtgcccaccc ctgcctacac cctgtggcag cactgccgtg gagttccctg gcagccaggg
     2761 caagcaggct tggcagtggt gcaaccctga ggacttcccg gaggtgttgt acccgcgaga
     2821 tgcgctggtg gtcgagatgc ccggagacag aggcgacggg acagagtcgc cccaggccgc
     2881 ccctgagtgc gccctggaca caaggcggcc cttggagact cagaggcaga ggcaggtgca
     2941 ggcactgtca gaggccaggc gcctgggcct ggctagggag gactgtcccc gtggtgacct
     3001 ggcccacgtg acactcccgc tgctcctggg aggtgtgacc cagggagcct ctgtacttga
     3061 cgatctttgg aggacccata agactgcgga gccgggaccg cccactttgg ggcaagaggc
     3121 cgaattcgcc actaacttct ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg
     3181 gccaatggtg agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt
     3241 caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
     3301 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
     3361 gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa
     3421 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
     3481 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
     3541 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     3601 cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga
     3661 cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga
     3721 cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa
     3781 cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     3841 cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtagctcga
     3901 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3961 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4021 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4081 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4141 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     4201 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4261 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4501 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4561 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4621 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4681 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4741 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4801 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4861 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4921 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4981 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     5041 tttcgcatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg
     5101 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac
     5161 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5221 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5281 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5341 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     5401 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5461 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     5521 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5581 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5641 gcctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5701 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5761 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     5821 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5881 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5941 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6001 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6061 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6121 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6181 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6241 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6301 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6901 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     6961 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7021 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7081 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7141 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7201 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7261 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7321 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7381 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7441 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7501 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7561 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7621 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7681 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7741 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7801 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7861 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7921 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7981 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8041 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8101 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     8161 tcccgatccc ctatggt
//
