LOCUS       V044293                 8150 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044293
VERSION     V044293
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..1780
                     /label="TMEM41B(NM_015012)"
                     /note="TMEM41B(NM_015012)"
                     /gene="TMEM41B"
     CDS             1823..2539
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             2549..2602
                     /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&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     polyA_signal    3717..3941
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3987..4415
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4429..4758
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4825..5619
                     /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    5793..5914
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5963..5979)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5987..6003
                     /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(6011..6041)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6056..6077
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6365..6950)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7121..7981)
                     /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(7982..8086)
                     /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 gcgaaaggca gagtcgccga acgatcgcag ttgggcgctc accacacgac
      961 ccccgtgggg gacggggcag cggggacgcg gggtctcgcg gcgcctggca gcagagacca
     1021 ccagaaggaa aaatcctggg tagaagctgg atcagcaaga atgtcactcc ttatattggt
     1081 gtccattttc ttatctgcag cttttgttat gtttttggta tataaaaatt ttcctcagct
     1141 tagtgaagaa gaaagagtga atatgaaggt tcccagagat atggatgatg ccaaggctct
     1201 aggaaaagtt ttatccaaat acaaggacac cttttatgtt caagtacttg tagcttattt
     1261 tgctacatat attttcttgc aaacatttgc tattccaggc tctatatttc tcagtatact
     1321 ctcagggttt ctttatccct ttccactagc cttatttctt gtttgtttgt gttctggact
     1381 tggtgcctct ttctgttata tgctttccta tttagttggg agaccagttg tatacaaata
     1441 cctaacagag aaagcagtaa aatggtcaca gcaggttgaa cgtcatagag aacatctcat
     1501 taactacatt atatttttga gaataacacc atttctgcct aattggttta ttaatatcac
     1561 atctcctgtg ataaacgtgc cattgaaagt tttttttatt ggtacttttc taggtgtcgc
     1621 acctccttct tttgtagcca ttaaggcagg aacaacactg tatcaactta caacagcagg
     1681 agaagctgtt tcctggaact caatatttat tctgatgatc ttggctgttc tttctattct
     1741 gccagccatc ttccaaaaaa aactaaagca gaaatttgag ctcgagtccg gactcagatc
     1801 tggcagcggt ggaggcgata tcatggtgag caagggcgag gagctgttca ccggggtggt
     1861 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1921 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1981 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2041 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2101 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2161 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2221 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2281 catggccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     2341 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2401 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2461 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2521 catggacgag ctgtacaagg gcagtggaga gggcagagga agtctgctaa catgcggtga
     2581 cgtcgaggag aatcctggcc caatgaggat atttgctgtc tttatattca tgacctactg
     2641 gcatttgctg aacgcattta ctgtcacggt tcccaaggac ctatatgtgg tagagtatgg
     2701 tagcaatatg acaattgaat gcaaattccc agtagaaaaa caattagacc tggctgcact
     2761 aattgtctat tgggaaatgg aggataagaa cattattcaa tttgtgcatg gagaggaaga
     2821 cctgaaggtt cagcatagta gctacagaca gagggcccgg ctgttgaagg accagctctc
     2881 cctgggaaat gctgcacttc agatcacaga tgtgaaattg caggatgcag gggtgtaccg
     2941 ctgcatgatc agctatggtg gtgccgacta caagcgaatt actgtgaaag tcaatgcccc
     3001 atacaacaaa atcaaccaaa gaattttggt tgtggatcca gtcacctctg aacatgaact
     3061 gacatgtcag gctgagggct accccaaggc cgaagtcatc tggacaagca gtgaccatca
     3121 agtcctgagt ggtaagacca ccaccaccaa ttccaagaga gaggagaagc ttttcaatgt
     3181 gaccagcaca ctgagaatca acacaacaac taatgagatt ttctactgca cttttaggag
     3241 attagatcct gaggaaaacc atacagctga attggtcatc ccagaactac ctctggcaca
     3301 tcctccaaat gaaaggactc acttggtaat tctgggagcc atcttattat gccttggtgt
     3361 agcactgaca ttcatcttcc gtttaagaaa agggagaatg atggatgtga aaaaatgtgg
     3421 catccaagat acaaactcaa agaagcaaag tgatacacat ttggaggaga cgggatccgg
     3481 cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggca gtgctggtgg
     3541 tagtgctggt ggtagtgctg gtggctctag aaccttgaga gaaatcgagg aattgttgag
     3601 gaaaatcatt gaagacagcg tgcgcagtgt ggccgagctg gaggacattg agaagtggct
     3661 gaagaaaatc tagctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     3721 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3781 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3841 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3901 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3961 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4021 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4081 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4141 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4201 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4261 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4321 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4381 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4441 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4501 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4561 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4621 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4681 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4741 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4801 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     4861 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4921 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4981 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5041 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5101 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5161 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5221 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5281 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5341 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5401 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5461 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5521 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5581 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5641 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5701 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5761 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5821 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5881 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5941 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6001 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6061 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6121 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6181 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6241 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6301 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6361 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6421 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6481 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6541 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6601 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6661 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6721 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6781 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6841 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6901 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6961 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7021 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7081 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7141 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7201 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7261 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7321 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7381 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7441 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7501 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7561 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7621 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7681 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7741 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7801 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7861 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7921 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7981 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8041 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8101 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
