LOCUS       V037266                 6834 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037266
VERSION     V037266
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..1649
                     /label="NGF(NM_002506)"
                     /note="NGF(NM_002506)"
                     /gene="NGF"
     CDS             1680..2393
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of red fluorescent protein
                     from Entacmaea quadricolor&nbsp;(Merzlyak et al., 2007)"
                     /transl_table=1
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN*"
                     /label="TagRFP"
     polyA_signal    2434..2658
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2704..3132
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3146..3475
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3542..4336
                     /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    4510..4631
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4680..4696)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4704..4720
                     /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(4728..4758)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4773..4794
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5082..5667)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5838..6698)
                     /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(6699..6803)
                     /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 ccatgttgtt ctacactctg atcacagctt
      961 ttctgatcgg catacaggcg gaaccacact cagagagcaa tgtccctgca ggacacacca
     1021 tcccccaagc ccactggact aaacttcagc attcccttga cactgccctt cgcagagccc
     1081 gcagcgcccc ggcagcggcg atagctgcac gcgtggcggg gcagacccgc aacattactg
     1141 tggaccccag gctgtttaaa aagcggcgac tccgttcacc ccgtgtgctg tttagcaccc
     1201 agcctccccg tgaagctgca gacactcagg atctggactt cgaggtcggt ggtgctgccc
     1261 ccttcaacag gactcacagg agcaagcggt catcatccca tcccatcttc cacaggggcg
     1321 aattctcggt gtgtgacagt gtcagcgtgt gggttgggga taagaccacc gccacagaca
     1381 tcaagggcaa ggaggtgatg gtgttgggag aggtgaacat taacaacagt gtattcaaac
     1441 agtacttttt tgagaccaag tgccgggacc caaatcccgt tgacagcggg tgccggggca
     1501 ttgactcaaa gcactggaac tcatattgta ccacgactca cacctttgtc aaggcgctga
     1561 ccatggatgg caagcaggct gcctggcggt ttatccggat agatacggcc tgtgtgtgtg
     1621 tgctcagcag gaaggctgtg agaagagccg gaggaggcgg atcaggaggc ggtggatcta
     1681 tggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
     1741 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     1801 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     1861 tcctggctac cagcttcatg tacggcagca gaaccttcat caaccacacc cagggcatcc
     1921 ccgacttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     1981 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     2041 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
     2101 cactcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc ctggaaggca
     2161 gaagcgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttcaagacca
     2221 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggacc
     2281 acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
     2341 ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat tgagactcta
     2401 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2461 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     2521 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     2581 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     2641 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     2701 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2761 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2821 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2881 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2941 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3001 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3061 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3121 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3181 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3241 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3301 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3361 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3421 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3481 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     3541 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3601 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3661 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3721 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3781 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3841 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3901 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3961 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4021 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4081 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4141 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4201 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4261 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4321 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4381 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4441 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4501 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     4561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     4621 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     4681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4861 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4921 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4981 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5041 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5101 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5161 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5221 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5281 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5341 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5401 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5461 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5521 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5581 agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag
     5641 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5701 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5761 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5821 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5881 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5941 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6001 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6061 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6121 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6181 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6241 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6301 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6361 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6421 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6481 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6541 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6601 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6661 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6721 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6781 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//
