LOCUS       V037658                 7270 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037658
VERSION     V037658
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             927..2075
                     /label="Rab3il1(NM_144538)"
                     /note="Rab3il1(NM_144538)"
                     /gene="Rab3il1"
     CDS             2113..2832
                     /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
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     polyA_signal    2870..3094
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3140..3568
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3582..3911
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3978..4772
                     /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    4946..5067
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5116..5132)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5140..5156
                     /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(5164..5194)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5209..5230
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5518..6103)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6274..7134)
                     /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(7135..7239)
                     /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 gtttaaagcc actgggatcc gccaccatgt ggagcggcca accccaacaa gacgagggcc
      961 tccctgtggg cctctctgct atcccagtcc cctggaaaaa cctaggcccc agcaaaagta
     1021 acagggagtc ctcaggaggc ctggtagaag cctccacttc ctgggaggag gcccagggtg
     1081 aggaacaccc ggcacctgct ccactggatg tgtcacgcct gcgcagttcc tccatggaga
     1141 tccgagagaa gggctccgag ttcctaaagg aggagctata caaagcccag aaggagctga
     1201 agctaaagga tgaagagtgt gaacggcttt gcaaagtacg tgcacagctg gagcaggagc
     1261 ttgaggagct gacagccagc ctgtttgagg aagctcacaa gatggttcgg gaagccaaca
     1321 tgaagcaggc gacatcggaa aagcagttga aggaggcttg gggcaagatc gacatgctac
     1381 aggcagaggt gacagccttg aagacattgg tcatcacatc cacaccagcc tctcccaacc
     1441 gtgagctcca cccacagctg ctgagcccca ccaaagctgg accccgaaag ggccactcac
     1501 gtcaaaagag caccagcagc tccctctgcc ctgttgtgtg ccccactgca gggcatatcc
     1561 ctacccctga caaagaaggc aaggaggtgg atacgacgct gtttgcagag ttccaagctt
     1621 ggagagcttc acctaccctg gataagagct gccccttcct agaaagggtg taccgggagg
     1681 acgtgggccc ttgccttgac ttcacagtgc aggagctctc agctctggtt cggactgctg
     1741 tggaggacaa cacactcacc attgagcctg tggcttcaca gacactgccc gctgtgaagg
     1801 tgcccactgt tgagtgtaac aacaccaaca catgtgccct gagtggcctg gcacgtacct
     1861 gccaccatcg aatccgcctg ggggactctg acagccacta ttacatctca ccgtcctccc
     1921 gggccaggat caccgcagta tgcaacttct tcacctatat tcgctacatc cagcaaggtc
     1981 tggttcggca ggatgctgag ccgatgttct gggagatcat gaggcttcgg aagggcatgt
     2041 cactagccaa gcttggcttc ttcccccagg aggccctcga gtggcggtgg aggaagcggc
     2101 ggaggtggat ctatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2161 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2221 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2281 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2341 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2401 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     2461 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     2521 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     2581 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     2641 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2701 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2761 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2821 ctgtacaagt agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2881 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2941 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3001 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3061 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3121 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3181 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3241 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3301 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3361 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3421 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3481 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3541 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     3601 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3661 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3781 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3841 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3901 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     3961 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4021 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4081 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4141 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4201 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4261 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4321 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4381 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4441 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4501 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4561 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4621 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4681 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4741 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4801 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4861 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4921 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4981 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5041 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5101 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5161 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5221 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5341 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5401 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5461 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5521 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5581 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5641 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5701 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5761 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5821 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5881 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5941 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6001 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtttttt
     6061 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6121 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6181 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6241 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6301 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6361 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6421 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6481 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6541 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6601 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6661 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6721 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6781 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6841 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6901 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6961 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     7021 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7081 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7141 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7201 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7261 acctgacgtc
//
