LOCUS       V030307                 8017 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030307
VERSION     V030307
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             931..1680
                     /codon_start=1
                     /note="more active than the original APEX due to the A134P
                     mutation"
                     /product="soybean ascorbate peroxidase, engineered to serve
                     as a monomeric tag for electron microscopy and live-cell
                     proteomics (Lam et al., 2015)"
                     /transl_table=1
                     /translation="MGKSYPTVSADYQDAVEKAKKKLRGFIAEKRCAPLMLRLAFHSAG
                     TFDKGTKTGGPFGTIKHPAELAHSANNGLDIAVRLLEPLKAEFPILSYADFYQLAGVVA
                     VEVTGGPKVPFHPGREDKPEPPPEGRLPDPTKGSDHLRDVFGKAMGLTDQDIVALSGGH
                     TIGAAHKERSGFEGPWTSNPLIFDNSYFTELLSGEKEGLLQLPSDKALLSDPVFRPLVD
                     KYAADEDAFFADYAEAHQKLSELGFADA"
                     /label="APEX2"
     CDS             1717..3537
                     /label="Atg16l1(NM_001108809)"
                     /note="Atg16l1(NM_001108809)"
                     /gene="Atg16l1"
     polyA_signal    3584..3808
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3854..4282
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4296..4625
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4692..5486
                     /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    5660..5781
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5830..5846)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5854..5870
                     /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(5878..5908)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5923..5944
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6232..6817)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6988..7848)
                     /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(7849..7953)
                     /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 cactagtcca gtgtggtgga attcgccacc atgggaaagt cttacccaac tgtgagtgct
      961 gattaccagg acgccgttga gaaggcgaag aagaagctca gaggcttcat cgctgagaag
     1021 agatgcgctc ctctaatgct ccgtttggca ttccactctg ctggaacctt tgacaagggc
     1081 acgaagaccg gtggaccctt cggaaccatc aagcaccctg ccgaactggc tcacagcgct
     1141 aacaacggtc ttgacatcgc tgttaggctt ttggagccac tcaaggcgga gttccctatt
     1201 ttgagctacg ccgatttcta ccagttggct ggcgttgttg ccgttgaggt cacgggtgga
     1261 cctaaggttc cattccaccc tggaagagag gacaagcctg agccaccacc agagggtcgc
     1321 ttgcccgatc ccactaaggg ttctgaccat ttgagagatg tgtttggcaa agctatgggg
     1381 cttactgacc aagatatcgt tgctctatct gggggtcaca ctattggagc tgcacacaag
     1441 gagcgttctg gatttgaggg tccctggacc tctaatcctc ttattttcga caactcatac
     1501 ttcacggagt tgttgagtgg tgagaaggaa ggtctccttc agctaccttc tgacaaggct
     1561 cttttgtctg accctgtatt ccgccctctc gttgacaaat atgcagcgga cgaagatgcc
     1621 ttctttgctg attacgctga ggctcaccaa aagctttccg agcttgggtt tgctgatgcc
     1681 ggtggaggcg gatcaggagg cggtggatct gatatcatgt cgtcgggcct ccgcgccgca
     1741 gactttcccc gctggaagcg ccacatcgcg gaggaactga ggcgccggga ccggctgcag
     1801 aggcaggcgt tcgaggagat cattctgcag tataccaagt tgctggagaa gtcagatctt
     1861 cattcagtat tgacccagaa actacaggct gaaaagcacg acgtaccaaa taggcatgaa
     1921 ataagtcctg ggcacgatgg tgcatggaat gacagtcagc tacaagaaat ggcccagttg
     1981 aagatgaaac accaggaaga gctgactgaa ctgcacaaga agcgtgggga gttagctcag
     2041 ttggtgattg acctgaataa ccagatgcag cagaaggaca aggagataca gacgaatgaa
     2101 gcaaagattg cagagtgttt acagaccatc tctgacctgg aggcagactg cctggacctg
     2161 cgcaccaaac tgcaggacct tgaggtagcc aaccagaccc tgaaggatga gtatgatgcc
     2221 ctgcagatta cttttactgc cctagaagag aaactgagga aaactactga ggagaaccag
     2281 gaactggtca ccaggtggat ggctgagaag gcccaagaag ccaatcgcct taatgcggag
     2341 aatgagaagg actccaggag gcgacaagca cggttgcaga aggagcttgc agaagcagca
     2401 aaggaacctc tacctgtcga acaggatgat gacattgaag tcattgtaga tgagacctca
     2461 gatcacacag aagagacctc tcctgtgcga gccatcagca gagcagctac taagcgactc
     2521 tcgcagcctg ctggaggcct tctggattct atcactaata tctttgggag acgctctgtc
     2581 tcttccatcc cagtccccca agatgtcgtg gacactcatc ctgcttctgg taaagatgtg
     2641 agagtcccaa ctactgcctc gtatgtcttt gatgcacatg acggagaggt caatgcagta
     2701 cagttcagtc caggctcccg gttactggcc actggaggca tggaccgcag ggttaaactt
     2761 tgggaagcat tcggagataa atgtgaattc aagggctccc tgtctggcag caatgcggga
     2821 attacaagca ttgaatttga tagtgctgga gcttacctat tagcagcttc aaatgatttt
     2881 gcaagccgaa tctggactgt ggatgattat cgattacggc acacacttac cggccacagc
     2941 gggaaagttc tctctgccaa gttcctgctg gacaatgcac ggatagtctc aggaagtcat
     3001 gaccggaccc tcaaactctg ggatctccgc agcaaagtct gtatcaaaac agtgttcgca
     3061 gggtccagct gcaatgacat tgtttgcact gagcaatgtg taatgagtgg acattttgac
     3121 aagaaaattc gtttctggga tatccggtca gagagtgtgg tccgagagat ggaactgtta
     3181 gggaagatca ctgctctgga cttaaacccc gagaggactg agctcctgag ctgctcccgt
     3241 gatgacctgc taaaaatcat tgacctgcgg acaaacgcca tcaagcagtc attcagtgca
     3301 cctggattca agtgtggctc tgactggacc cgagttgtct tcagccctga tggcagttac
     3361 gtggcagcag gctcagctga cggctctctt tatgtatgga gtgtgctcac ggggaaagta
     3421 gagaaggttc tctccaagca gcacagctct tctatcaatg cggtggcgtg ggccccctct
     3481 ggcttacatg ttgtcagtgt ggacaaagga agcagagctg tgctatgggc acagccttga
     3541 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3601 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3661 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3721 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3781 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3841 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4201 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4621 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5521 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5581 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5641 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5701 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5761 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5821 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5881 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5941 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6001 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6061 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6121 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6181 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6241 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6301 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6361 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6421 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6481 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6541 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6601 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6661 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6721 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6781 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6841 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6901 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6961 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7021 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7081 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7141 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7201 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7261 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7321 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7381 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7441 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7501 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7561 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7621 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7681 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7741 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7801 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7861 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7921 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7981 cgtcgacgga tcgggagatc tcccgatccc ctatggt
//
