LOCUS       V033862                 8002 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033862
VERSION     V033862
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3492
                     /label="Afap1l2(NM_001177796)"
                     /note="Afap1l2(NM_001177796)"
                     /gene="Afap1l2"
     CDS             3499..3564
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3602..3826
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3872..4300
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4314..4643
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4710..5504
                     /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    5678..5799
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5848..5864)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5872..5888
                     /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(5896..5926)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5941..5962
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6250..6835)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7006..7866)
                     /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(7867..7971)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggagc gctacaaagc acagggctgt tgctgtctgg tggtgcagag aagaatcctg
     1021 caggtctctg cttccctgga gcagctgctg acagagctgg acgacttcct taaggtcctc
     1081 gaccaggaga acctgagcag tgcagccgtg ctgaagaaga gcggcctgtc agagctcctg
     1141 cggctttaca ccaagagcag cagttctgat gaggagtata tttacatgaa caaagtgtca
     1201 gtcaacgggg aacaaaactc agcatctcca gacaaagttc ccgaggagca gggtccactg
     1261 accaatgggg agcccagcca gcattcctca gcccctcaga agagccttcc agacctccct
     1321 ccgcccaaga tgattccaga gaggaaacag cccaccgttc caaagatcga gtcccctgag
     1381 ggctactatg aagaggctga gccatttgac agatccatca atgaggatgg agaggctgtg
     1441 agcagctcct acgagtccta cgatgaggac gagaacagca aaggcaaggc tgccccctac
     1501 cagtggccct cgcccgaggc cagcattgag ctgatgcgtg atgcccgcat ctgtgccttc
     1561 ctgtggcgca agaagtggct aggccagtgg gccaagcagc tctgtgtgat cagggacacc
     1621 aggctcttgt gctacaaatc gtccaaagac cacagtcctc agctggacgt gaacctgcgg
     1681 ggcagcagcg tcgtgcacaa ggagaagcaa gtgcggaaga aaggacacaa gctcaagatc
     1741 acaccgatga acgctgacgt tatcgtgctg ggcctgcaga gcaaggacca agctgagcag
     1801 tggcttcggg tcatccagga ggtgagtggc ctgccttctg aaggagcctc agagggaaac
     1861 cagtacacac cagatgccca gcgcctcaac tgtcagaaac cagacatagc tgagaagtac
     1921 ctgtcagcag cggagtacgg gatcaccatc aacggccacc ctgagatccc agagaccaaa
     1981 gatgtcaaga agaaatgctc tgctggcctc aagctgagca accttatgaa cctgggcagg
     2041 aagaaatcta cctcgctgga gcccccggag agatcccttg agacatccag ctacctgaat
     2101 gtgctggtga acagtcaatg gaagtcacgc tggtgcttcg ttagagacag ccacctgcac
     2161 ttctaccagg accggaaccg gagcaaggtg gcccagcagc ccctcagcct ggtgggctgt
     2221 gacgtgcttc cagatcctag ccccgaccac ctgtattcct tccgtattct ccacaacggc
     2281 gaggagctag ccaagcttga ggccaagtct tcagaggaga tgggccactg gctaggcctc
     2341 ctgctctcag agtcgggctc caagactgac ccggaagagc tcacctacga ctatgtggat
     2401 gctgaaaggg tttcctgtat tgtgagtgcg gccaaaacct ctctcttact gatgcagaga
     2461 aagttctcag agcccaacac atacatcgat ggcctgccca gccgggattg ccaggacgat
     2521 ctgtatgatg atgtggaggt atcagagctg atagccgtgg tggagcctgc cgaggaagcc
     2581 gcccctgctg tcgatgctaa cagtggtagt gagccagaca gagtctacct ggatctcaca
     2641 ccggtcaagt ccttcctgca cagtagcagt gaggctcagg cgcaggcctc tctcccagca
     2701 gtgccacatc aggacgatgt agctgagacc ctcacagtag acccaaagcc aggcaccacc
     2761 ccagaggagc cccacacaga gtctccagga gacccagagg tgcagcagag gcaaccagag
     2821 gttcaagagt cttcagagcc tatcgagccc accccgcgaa tcaccatggt taagctacag
     2881 gccgagcagc agagaatctc cttcccagcc aactgcccag acaccatggc ttctgccccc
     2941 atcgctgcca gcccacctgt gaaggagaag ctgagagtga ccagtgcaga gatcaaactt
     3001 gggaagaatc ggacagaggc ggaagtaaag cggtacacgg aggagaagga gaggctggag
     3061 aggagcaagg aggagatccg agggcacctg gctcagctcc gcagagagaa gcgggagctc
     3121 aaagagaccc tgttgagatg cacagataag ggggtcctgg ccaagctgga gcagacactg
     3181 aagaaaatag acgaggaatg ccggatggag gagagcaggc gtgtggacct tgagctcagc
     3241 atcatggagg tgaaggacaa cctgaagaag gcagaggctg ggcccgtgac cctgggcacc
     3301 actgtggata ccacgcacct ggacaacatg agccctcgtc cacagcccaa agctgccacc
     3361 cccaaccccc caccagactc cacgccagtc aactctgcat ctgtgctcaa gaacaggcct
     3421 ctttccgtca tggtcacagg caaaggcact gtcctgcaga aagccaagga atgggagaag
     3481 aaaggagcca gtctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc
     3541 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg
     3601 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3661 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3721 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3781 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3841 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3901 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3961 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4021 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4081 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4141 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4201 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4261 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4321 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4381 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4441 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4501 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4561 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4621 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4681 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4741 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4801 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4861 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4921 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4981 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5041 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5101 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5161 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5221 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5281 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5341 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5401 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5461 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5521 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5581 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5641 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5701 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5761 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5821 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5881 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5941 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6001 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6061 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6661 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6781 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7141 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7981 ccgaaaagtg ccacctgacg tc
//
