LOCUS       V045770                 7228 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045770
VERSION     V045770
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             931..1863
                     /codon_start=1
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /product="near-infrared fluorescent protein&nbsp;with an
                     emission peak at 670 nm, engineered from a bacterial
                     phytochrome&nbsp;(Shcherbakova and Verkhusha, 2013)"
                     /protein_id=""
                     /transl_table=1
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
                     /label="iRFP670"
     CDS             1900..2664
                     /label="YWHAE(NM_006761)"
                     /note="YWHAE(NM_006761)"
                     /gene="YWHAE"
     CDS             2671..2760
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    2795..3019
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3065..3493
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3507..3836
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3903..4697
                     /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    4871..4992
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5041..5057)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5065..5081
                     /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(5089..5119)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5134..5155
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5443..6028)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6199..7059)
                     /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(7060..7164)
                     /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 ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atggcgcgta aggtcgatct cacctcctgc
      961 gatcgcgagc cgatccacat ccccggcagc attcagccgt gcggctgcct gctagcctgc
     1021 gacgcgcagg cggtgcggat cacgcgcatt acggaaaatg ccggcgcgtt ctttggacgc
     1081 gaaactccgc gggtcggtga gctactcgcc gattacttcg gcgagaccga agcccatgcg
     1141 ctgcgcaacg cactggcgca gtcctccgat ccaaagcgac cggcgctgat cttcggttgg
     1201 cgcgacggcc tgaccggccg caccttcgac atctcactgc atcgccatga cggtacatcg
     1261 atcatcgagt tcgagcctgc ggcggccgaa caggccgaca atccgctgcg gctgacgcgg
     1321 cagatcatcg cgcgcaccaa agaactgaag tcgctcgaag agatggccgc acgggtgccg
     1381 cgctatctgc aggcgatgct cggctatcac cgcgtgatgt tgtaccgctt cgcggacgac
     1441 ggctccggga tggtgatcgg cgaggcgaag cgcagcgacc tcgagagctt tctcggtcag
     1501 cactttccgg cgtcgctggt cccgcagcag gcgcggctac tgtacttgaa gaacgcgatc
     1561 cgcgtggtct cggattcgcg cggcatcagc agccggatcg tgcccgagca cgacgcctcc
     1621 ggcgccgcgc tcgatctgtc gttcgcgcac ctgcgcagca tctcgccctg ccatctcgaa
     1681 tttctgcgga acatgggcgt cagcgcctcg atgtcgctgt cgatcatcat tgacggcacg
     1741 ctatggggat tgatcatctg tcatcattac gagccgcgtg ccgtgccgat ggcgcagcgc
     1801 gtcgcggccg aaatgttcgc cgacttctta tcgctgcact tcaccgccgc ccaccaccaa
     1861 cgcggtggag gcggatcagg aggcggtgga tctctcgaga tggatgatcg agaggatctg
     1921 gtgtaccagg cgaagctggc cgagcaggct gagcgatacg acgaaatggt ggagtcaatg
     1981 aagaaagtag cagggatgga tgtggagctg acagttgaag aaagaaacct cctatctgtt
     2041 gcatataaga atgtgattgg agctagaaga gcctcctgga gaataatcag cagcattgaa
     2101 cagaaagaag aaaacaaggg aggagaagac aagctaaaaa tgattcggga atatcggcaa
     2161 atggttgaga ctgagctaaa gttaatctgt tgtgacattc tggatgtact ggacaaacac
     2221 ctcattccag cagctaacac tggcgagtcc aaggttttct attataaaat gaaaggggac
     2281 taccacaggt atctggcaga atttgccaca ggaaacgaca ggaaggaggc tgcggagaac
     2341 agcctagtgg cttataaagc tgctagtgat attgcaatga cagaacttcc accaacgcat
     2401 cctattcgct taggtcttgc tctcaatttt tccgtattct actacgaaat tcttaattcc
     2461 cctgaccgtg cctgcaggtt ggcaaaagca gcttttgatg atgcaattgc agaactggat
     2521 acgctgagtg aagaaagcta taaggactct acacttatca tgcagttgtt acgtgataat
     2581 ctgacactat ggacttcaga catgcagggt gacggtgaag agcagaataa agaagcgctg
     2641 caggacgtgg aagacgaaaa tcagctcgag gaacaaaaac taatatcaga ggaagatttg
     2701 gaacagaaac taatcagtga agaagattta gagcaaaagt taatttccga ggaggactta
     2761 tgagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2821 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     2881 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     2941 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3001 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3061 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3121 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3181 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3241 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3301 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3361 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3421 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3481 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3541 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3601 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3661 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3721 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3781 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3841 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     3901 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3961 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4021 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4081 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4141 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4201 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4261 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4321 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4381 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4441 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4501 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4561 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4621 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4681 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4741 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4801 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4861 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4921 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4981 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5041 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5101 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5161 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5221 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5281 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5341 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5401 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5461 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5521 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5581 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5641 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5701 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5761 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5821 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5881 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5941 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca
     6001 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6061 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6121 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6181 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6241 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6301 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6361 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6421 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6481 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6541 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6601 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6661 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6721 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6781 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6841 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6901 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6961 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7021 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7081 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7141 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7201 atcgggagat ctcccgatcc cctatggt
//
