LOCUS       V024187                 8883 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024187
VERSION     V024187
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..1600
                     /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"
     CDS             1622..4372
                     /label="Hk2(NM_013820)"
                     /note="Hk2(NM_013820)"
                     /gene="Hk2"
     polyA_signal    4450..4674
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4720..5148
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5162..5491
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5558..6352
                     /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    6526..6647
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6696..6712)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6720..6736
                     /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(6744..6774)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6789..6810
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7098..7683)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7854..8714)
                     /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(8715..8819)
                     /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 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      961 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1021 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1081 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1141 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1201 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1261 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1321 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1381 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1441 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1501 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1561 cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca gatctcgagc
     1621 tatgatcgcc tcgcatatga tcgcctgctt attcacggag ctcaaccaaa accaagtgca
     1681 gaaggttgac cagtatctct accacatgcg tctctcagat gagacccttc tggagatttc
     1741 taggcggttc cggaaggaga tggagaaggg gctaggagct accacacacc ctacagcagc
     1801 tgtgaagatg ctgcccacct ttgtgaggtc aactccggat gggacagaac atggcgagtt
     1861 cctggctctg gaccttggag gaaccaactt ccgtgtgctc cgagtaaggg tgacagacaa
     1921 tggtctccag agggtggaga tggagaacca gatctacgcc attcccgagg acatcatgcg
     1981 gggcagtgga acccagctgt ttgaccacat tgccgaatgc ctggccaact tcatggacaa
     2041 gctacagatc aaagagaaga agctcccgct gggtttcacc ttctcgttcc cctgccacca
     2101 gacgaaactg gatgagagtt tcttggtctc atggactaag gggttcaagt ccagtggcgt
     2161 ggaaggcaga gatgtggtgg atctgatccg gaaggccatc cagcgcagag gggactttga
     2221 cattgacatt gtggctgtgg tgaatgacac ggttgggacc atgatgactt gtggttatga
     2281 tgatcagaac tgcgagattg gtctcattgt gggtactggc agtaatgcct gctacatgga
     2341 ggagatgcgt cacattgaca tggtggaggg cgacgaggga cggatgtgca tcaacatgga
     2401 gtggggggcc tttggggacg acggtacact caatgacatc cgaactgagt ttgacagaga
     2461 gatcgacatg ggctcactga accctgggaa gcagctgttt gagaagatga tcagcgggat
     2521 gtacatgggg gagctggtca gactcatcct ggtgaagatg gccaaggccg agctgctgtt
     2581 ccaagggaaa ctcagcccag agctccttac cactggctcc ttcgagacca aagatgtctc
     2641 ggatattgaa gacgataagg acggaattca gaaggcctac cagatcctgg tgcgcctggg
     2701 tctgagcccg ctgcaggagg actgtgtggc cacgcaccga atctgccaga ttgtgtccac
     2761 gcgctcagcc agcctgtgtg cagccaccct ggccgcggtg ctgtggcgga tcaaagagaa
     2821 caagggcgag gagcgactcc gctccaccat cggcgtggat ggctctgtct acaagaaaca
     2881 tccccatttt gccaagcgtc tccataaggc agtgaggagg ctggtgcccg attgtgatgt
     2941 ccgcttcctc cgctctgagg atggcagcgg caagggggcg gctatggtga cggcggtggc
     3001 ttaccgtctg gctgaccaac accgggcccg ccagaagacc ctggagtctc tgaagctgag
     3061 ccatgaacag ctgctggagg ttaagagaag gatgaaggtg gaaatggagc agggtctgag
     3121 caaggagacg catgaggccg cccctgtgaa gatgttgccc acttatgtgt gtgccactcc
     3181 agacggtaca gagaaaggag acttcctggc cttggatctt ggaggaacaa acttccgggt
     3241 cctgctggtg cgcgtgcgta atggcaagcg aaggggcgta gagatgcata acaagatcta
     3301 ctccatccca caggaagtca tgcatggcac tggagaagag cttttcgacc acattgtcca
     3361 gtgcatcgcg gacttccttg agtacatggg catgaagggc gtgtccctac ctttgggttt
     3421 caccttctcc ttcccttgcc agcagaacag cctggaccag agcatcctcc tcaagtggac
     3481 aaagggattc aaggcatccg gctgcgaggg tgaagatgtg gtcaccttgc tgaaggaagc
     3541 cattcgccgg cgagaggagt ttgacctgga tgtggtggcc gtggtaaatg acacagttgg
     3601 gactatgatg acttgtggct atgaagaccc tcactgtgaa gttggcctca ttgttggcac
     3661 tggaagcaat gcctgctaca tggaggagat gcgtaatgtg gaactggtgg acggagagga
     3721 ggggcggatg tgtgtcaaca tggagtgggg agcgttcggg gacaacggct gcctggatga
     3781 cctgcgcaca gtgtttgacg tcgctgtgga cgagctctct ctcaaccctg gcaaacagag
     3841 gtttgagaag atgatcagcg gcatgtactt gggcgagatt gtgcgcaaca tcctgatcga
     3901 tttcacaaag cgggggctgc tcttccgtgg ccgcatctca gagcgcctca agacaagggg
     3961 aatcttcgaa actaagttcc tgtctcagat cgagagtgac tgcctggccc tgctacaggt
     4021 ccgagccatc ttgcgccacc tagggctgga gagcacgtgt gacgacagca tcattgtgaa
     4081 ggaggtgtgc actgtggtcg cccggcgcgc ggcccagctc tgtggcgcag gcatggccgc
     4141 cgtggtggac aagataagag agaaccgtgg actggacaac ctcaaagtga cggtgggcgt
     4201 ggacgggaca ctgtacaagc ttcatcctca ctttgccaaa gtcatgcatg agacggtgag
     4261 agatctggct ccgaaatgtg acgtgtcctt cctggaatcc gaggacggca gtgggaaggg
     4321 agcggctctc atcactgctg tggcctgtcg catccgggag gccgggcaga gataggaatt
     4381 ctgcagtcga cggtaccgcg ggcccgggat cctctagagg gcccgtttaa acccgctgat
     4441 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4501 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4561 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4621 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4681 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4741 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4801 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4861 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4921 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4981 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5041 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5101 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     5161 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     5221 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     5281 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5341 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5401 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     5461 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     5521 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     5581 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     5641 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     5701 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     5761 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     5821 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     5881 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     5941 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     6001 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     6061 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     6121 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     6181 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     6241 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     6301 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     6361 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6421 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6481 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6541 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6601 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6661 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6721 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6781 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6841 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     7021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7621 accgctggta gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7981 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8161 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8821 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8881 ggt
//
