LOCUS       V026773                 8108 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V026773
VERSION     V026773
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..1591
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             1649..2893
                     /label="Lamp2(NM_001017959)"
                     /note="Lamp2(NM_001017959)"
                     /gene="Lamp2"
     polyA_signal    3675..3899
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3945..4373
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4387..4716
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4783..5577
                     /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    5751..5872
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5921..5937)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5945..5961
                     /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(5969..5999)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6014..6035
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6323..6908)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7079..7939)
                     /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(7940..8044)
                     /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 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
      961 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     1021 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     1081 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     1141 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     1201 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     1261 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     1321 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     1381 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     1441 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     1501 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     1561 ctccaccggc ggcatggacg agctgtacaa gtccggactc agatctcgag ctggaggtgg
     1621 aggtagtgga ggaggcggaa gcgaattcat gtgcctctct ccggttaaag gcgcaaagct
     1681 catcctgatc tttctgttcc taggagccgt tcagtccaat gcattgatag ttaatttgac
     1741 agattcaaag ggtacttgcc tttatgcaga atgggagatg aatttcacaa taacatatga
     1801 aactacaaac caaaccaata aaactataac cattgcagta cctgacaagg cgacacacga
     1861 tggaagcagt tgtggggatg accggaatag tgccaaaata atgatacaat ttggattcgc
     1921 tgtctcttgg gctgtgaatt ttaccaagga agcatctcat tattcaattc atgacatcgt
     1981 gctttcctac aacactagtg atagcacagt atttcctggt gctgtagcta aaggagttca
     2041 tactgttaaa aatcctgaga atttcaaagt tccattggat gtcatcttta agtgcaatag
     2101 tgttttaact tacaacctga ctcctgtcgt tcagaaatat tggggtattc acctgcaagc
     2161 ttttgtccaa aatggtacag tgagtaaaaa tgaacaagtg tgtgaagaag accaaactcc
     2221 caccactgtg gcacccatca ttcacaccac tgccccgtcg actacaacta cactcactcc
     2281 aacttcaaca cccactccaa ctccaactcc aactccaacc gttggaaact acagcattag
     2341 aaatggcaat actacctgtc tgctggctac catggggctg cagctgaaca tcactgagga
     2401 gaaggtgcct ttcattttta acatcaaccc tgccacaacc aacttcaccg gcagctgtca
     2461 acctcaaagt gctcaactta ggctgaacaa cagccaaatt aagtatcttg actttatctt
     2521 tgctgtgaaa aatgaaaaac ggttctatct gaaggaagtg aatgtctaca tgtatttggc
     2581 taatggctca gctttcaaca tttccaacaa gaaccttagc ttctgggatg cccctctggg
     2641 aagttcttat atgtgcaaca aagagcaggt gctttctgtg tctagagcgt ttcagatcaa
     2701 cacctttaac ctaaaggtgc aaccttttaa tgtgacaaaa ggacagtatt ctacagctca
     2761 agactgcagt gcagatgaag acaacttcct tgtgcccata gcggtgggag cagctctggg
     2821 aggagtactt attctagtgt tgctggctta ttttattggt ctcaagcgcc atcatactgg
     2881 atatgagcaa tttggtggag gcggatcagg aggcggtgga tctatgagca cagaaagcat
     2941 gatccgcgac gtggaactgg cagaagaggc actcccccaa aagatggggg gcttccagaa
     3001 ctccaggcgg tgcctatgtc tcagcctctt ctcattcctg cttgtggcag gggccaccac
     3061 gctcttctgt ctactgaact tcggggtgat cggtccccaa agggatgaga agttcccaaa
     3121 tggcctccct ctcatcagtt ctatggccca gaccctcaca ctcagatcat cttctcaaaa
     3181 ttcgagtgac aagcctgtag cccacgtcgt agcaaaccac caagtggagg agcagctgga
     3241 gtggctgagc cagcgcgcca acgccctcct ggccaacggc atggatctca aagacaacca
     3301 actagtggtg ccagccgatg ggttgtacct tgtctactcc caggttctct tcaagggaca
     3361 aggctgcccc gactacgtgc tcctcaccca caccgtcagc cgatttgcta tctcatacca
     3421 ggagaaagtc aacctcctct ctgccgtcaa gagcccctgc cccaaggaca cccctgaggg
     3481 ggctgagctc aaaccctggt atgagcccat atacctggga ggagtcttcc agctggagaa
     3541 gggggaccaa ctcagcgctg aggtcaatct gcccaagtac ttagactttg cggagtccgg
     3601 gcaggtctac tttggagtca ttgctctgtg aggatcctct agagggcccg tttaaacccg
     3661 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3721 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3781 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3841 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3901 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4141 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4261 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     4321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4381 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4441 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4501 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4561 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4621 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4681 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4741 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4801 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4861 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4921 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     4981 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     5041 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     5101 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     5161 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5221 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5281 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     5341 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5401 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5461 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5521 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5581 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5641 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5701 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5761 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5821 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5881 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5941 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     6001 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6061 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6121 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6181 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6241 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6301 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6361 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6421 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6481 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6541 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6601 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6661 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6721 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6781 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6841 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6901 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6961 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     7021 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     7081 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7141 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7201 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7261 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7321 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7381 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7441 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7501 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7561 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7621 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7681 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7741 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7801 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7861 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7921 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7981 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     8041 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     8101 cctatggt
//
