LOCUS       V037777                 8261 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037777
VERSION     V037777
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             884..1591
                     /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             1628..3781
                     /label="TARS2(NM_025150)"
                     /note="TARS2(NM_025150)"
                     /gene="TARS2"
     polyA_signal    3828..4052
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4098..4526
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4540..4869
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4936..5730
                     /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    5904..6025
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6074..6090)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6098..6114
                     /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(6122..6152)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6167..6188
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6476..7061)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7232..8092)
                     /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(8093..8197)
                     /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 ggctagcgct accggtcgcc accatggtga gcaagggcga
      901 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gggtggaggc ggatcaggag gcggtggatc
     1621 tctcgagatg gccctgtatc agaggtggcg gtgtctccgg ctccaaggtt tacaggcttg
     1681 caggctacac acggcagttg tgtcgacccc tccacgctgg ttggcagagc ggcttggcct
     1741 ttttgaggag ctgtgggctg ctcaggtaaa gagattagca agcatggcac agaaggaacc
     1801 ccggactatt aagatatcac ttcctggagg ccagaaaatt gatgctgtgg catggaacac
     1861 aaccccctac caactagccc ggcagatcag ttcaacactg gcagatactg cagtggctgc
     1921 tcaagtgaat ggagaacctt atgatctgga gcggcccttg gagacagatt ctgacctcag
     1981 atttctgaca ttcgattccc cagaggggaa agcagtgttc tggcactcca gcacccatgt
     2041 cctgggggca gcagctgaac aattcctagg tgctgttctc tgcagaggtc caagtacaga
     2101 atatggcttt taccatgatt tcttcctggg aaaggagagg acaatccggg gctcagagct
     2161 gcctgttttg gagcggattt gccaggaact tacagctgct gctcgaccct tccggaggct
     2221 agaggcttca cgggatcagc ttcgccagtt gttcaaggat aaccccttta agcttcactt
     2281 gattgaggag aaagtgacag gtccaacagc aacagtatat gggtgtggca cattggttga
     2341 cctttgccag ggcccccacc ttcggcatac tggacagatt ggaggactga agctgctatc
     2401 gaactcatca tccttatgga ggtcttcagg ggccccagag acactgcaga gagtgtcagg
     2461 gatttccttc cccacaacag aattgctgag ggtctgggaa gcatggaggg aggaagcaga
     2521 attgcgggac caccggcgca ttgggaagga acaggagctc ttcttcttcc atgaactgag
     2581 ccctgggagc tgcttcttcc tgccacgagg gacaagggtg tataatgcac tagtggcgtt
     2641 tatcagggct gagtatgccc atcgtggttt ctccgaggtg aaaactccca cactgttttc
     2701 tacgaagctc tgggaacagt cagggcactg ggagcattat caggaagaca tgtttgccgt
     2761 gcagccccca ggctctgaca ggcctcccag ctcccagagt gacgattcta ccaggcatat
     2821 cacagataca ctcgccctca agcctatgaa ctgccctgca cactgcctga tgttcgccca
     2881 ccggcccaga tcctggcggg aactgcccct gcgactagct gactttgggg ctctacaccg
     2941 ggccgaagcc tctggtggtc tggggggact gacccgactg cggtgcttcc agcaggatga
     3001 cgctcacatc ttctgtacaa cagatcagct ggaagcagag atccaaagct gtcttgattt
     3061 cctccgttcc gtctatgccg ttcttggctt ctccttccgc ctggcactgt ccacccggcc
     3121 atctggcttc ctgggggacc cttgcctttg ggaccaggcc gaacaggtcc ttaaacaggc
     3181 cctgaaggaa tttggagaac cctgggacct caactctgga gatggtgcct tctatggacc
     3241 taagattgac gtgcacctcc acgatgccct gggccggcca catcagtgtg ggacaattca
     3301 gcttgacttc caactgcccc tgagatttga cctccagtat aaggggcagg cgggtgccct
     3361 ggagcgtcca gtcctcattc accgagcagt gctcggttct gtggaaagac tgttgggagt
     3421 gctggcagaa agctgcgggg ggaaatggcc actgtggctg tccccgttcc aggtggtggt
     3481 catccctgtg gggagtgagc aagaggaata cgccaaagag gcacagcaga gcctgcgggc
     3541 tgcaggactg gtcagtgacc tggatgcaga ctctggactg accctcagcc ggagaatccg
     3601 ccgggcccag cttgcccact acaattttca gtttgtggtt ggccagaaag agcaaagtaa
     3661 gagaacagtg aacattcgga ctcgagataa tcgtcgcctt ggggagtggg acttgcctga
     3721 ggctgtgcag cgactggtgg agctacagaa cacgagggtc ccaaatgccg aagaaatttt
     3781 ctgaggatcc tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     3841 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     3901 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     3961 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     4021 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     4081 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4141 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4201 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4261 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4321 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4381 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4441 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     4501 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     4561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4621 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4681 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4741 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4801 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4861 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     4921 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     4981 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5041 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5101 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5161 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5221 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5281 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5341 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5401 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5461 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5521 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5581 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5641 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5701 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     5761 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     5821 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     5881 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     5941 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6001 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     6061 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     6121 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     6181 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     6241 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     6301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     6361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     6421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     6481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     6541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     6601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     6661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     6721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     7021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8221 ctgacgtcga cggatcggga gatctcccga tcccctatgg t
//
