LOCUS       pLenti-H2B-iRFP7        7760 bp    DNA     circular UNA 25-JUN-2019
DEFINITION  .
ACCESSION   urn.local...2d-amswjoj
VERSION     urn.local...2d-amswjoj
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     CDS             519..914
                     /codon_start=1
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYK
                     VLKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLL
                     LPGELAKHAVSEGTKAITKYTSAKDPPVAT"
                     /transl_table=1
                     /label="H2B"
     primer_bind     649..669
                     /Sequence="AGGTTCTGAAGCAGGTCCACC"
                     /Hairpin_Tm="31.4"
                     /Tm="62.0"
                     /Self_Dimer_Tm="20.4"
                     /%GC="57.1"
                     /created_by="primer3"
                     /label="pZM19.f"
     primer_bind     872..890
                     /Sequence="CATCACCAAGTACACCAGC"
                     /Hairpin_Tm="None"
                     /Tm="55.9"
                     /Self_Dimer_Tm="None"
                     /%GC="52.6"
                     /created_by="primer3"
                     /label="spZM1.f"
     primer_bind     complement(896..914)
                     /Sequence="GGTGGCGACCGGTGGATCC"
                     /Hairpin_Tm="30.6"
                     /Tm="65.2"
                     /Self_Dimer_Tm="14.1"
                     /%GC="73.7"
                     /created_by="primer3"
                     /label="pZM12.r"
     source          <915..>1862
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     CDS             <915..1862
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAG
                     SDNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAG
                     FIGSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVR
                     KITGFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRI
                     IPDINYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGE
                     RLWGLIVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
                     /label="iRFP720"
     primer_bind     1863..1890
                     /Sequence="GTACCTTTAAGACCAATGACTTACAAGG"
                     /Hairpin_Tm="39.4"
                     /Tm="60.1"
                     /Self_Dimer_Tm="20.7"
                     /%GC="39.3"
                     /created_by="primer3"
                     /modified_by="User"
                     /label="pZM11.f"
     misc_feature    complement(1875..1894)
                     /Sequence="GCTGCCTTGTAAGTCATTGG"
                     /Hairpin_Tm="None"
                     /Tm="57.1"
                     /Self_Dimer_Tm="None"
                     /%GC="50.0"
                     /created_by="primer3"
                     /label="spZM2.r"
                     /note="Geneious type: primer_bind_reverse"
     LTR             1933..2166
                     /note="<html><body>self-inactivating&nbsp;3' long terminal
                     repeat (LTR) from HIV-1</body></html>"
                     /label="3' LTR (ΔU3)"
     polyA_signal    2238..2359
                     /note="<html><body>SV40 polyadenylation
                     signal</body></html>"
                     /label="SV40 poly(A) signal"
     rep_origin      2399..2534
                     /note="<html><body>SV40 origin of
                     replication</body></html>"
                     /label="SV40 ori"
     promoter        complement(2555..2573)
                     /note="<html><body>promoter for bacteriophage T7 RNA
                     polymerase</body></html>"
                     /label="T7 promoter"
     primer_bind     complement(2583..2599)
                     /note="<html><body>common sequencing primer, one of
                     multiple similar variants</body></html>"
                     /label="M13 fwd"
     rep_origin      2741..3196
                     /note="<html><body>f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand
                     synthesis</body></html>"
                     /direction="RIGHT"
                     /label="f1 ori"
     promoter        3222..3326
                     /gene="<html><body><i>bla</i></body></html>"
                     /label="AmpR promoter"
     CDS             order(3327..3395,3396..4187)
                     /gene="<html><body><i>bla</i></body></html>"
                     /note="<html><body>confers resistance to ampicillin,
                     carbenicillin, and related antibiotics</body></html>"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVG
                     YIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLV
                     EYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTR
                     LDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGP
                     LLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQI
                     AEIGASLIKHW*"
                     /product="<html><body>β-lactamase</body></html>"
                     /transl_table=1
                     /label="AmpR"
     misc_feature    complement(3441..3461)
                     /Sequence="GATGTAACCCACTCGTGCACC"
                     /Hairpin_Tm="None"
                     /Tm="61.3"
                     /Self_Dimer_Tm="12.2"
                     /%GC="57.1"
                     /created_by="primer3"
                     /label="pZM20.r"
                     /note="Geneious type: primer_bind_reverse"
     rep_origin      4358..4946
                     /note="<html><body>high-copy-number ColE1/pMB1/pBR322/pUC
                     origin of replication</body></html>"
                     /direction="RIGHT"
                     /label="ori"
     protein_bind    5234..5255
                     /note="<html><body>CAP binding activates transcription in
                     the presence of cAMP.</body></html>"
                     /bound_moiety="<html><body><i>E. coli</i> catabolite
                     activator protein</body></html>"
                     /label="CAP binding site"
     promoter        order(5270..5275,5276..5293,5294..5300)
                     /note="<html><body>promoter for the <i>E. coli lac</i>
                     operon</body></html>"
                     /label="lac promoter"
     protein_bind    5308..5324
                     /note="<html><body>The <i>lac</i> repressor binds to the
                     <i>lac</i> operator to inhibit transcription in <i>E.
                     coli</i>. This inhibition can be relieved by adding
                     lactose or isopropyl-β-D-thiogalactopyranoside
                     (IPTG).</body></html>"
                     /bound_moiety="<html><body><i>lac</i> repressor encoded by
                     <i>lacI</i></body></html>"
                     /label="lac operator"
     primer_bind     5332..5348
                     /note="<html><body>common sequencing primer, one of
                     multiple similar variants</body></html>"
                     /label="M13 rev"
     promoter        5369..5387
                     /note="<html><body>promoter for bacteriophage T3 RNA
                     polymerase</body></html>"
                     /label="T3 promoter"
     promoter        5413..5641
                     /note="<html><body>Rous sarcoma virus
                     enhancer/promoter</body></html>"
                     /label="RSV promoter"
     LTR             5642..5822
                     /note="<html><body>truncated 5' long terminal repeat (LTR)
                     from HIV-1</body></html>"
                     /label="5' LTR (truncated)"
     misc_feature    5869..5994
                     /note="<html><body>packaging signal of human
                     immunodeficiency virus type 1</body></html>"
                     /label="HIV-1 Ψ"
     misc_feature    6487..6720
                     /note="<html><body>The Rev response element (RRE) of HIV-1
                     allows for Rev-dependent mRNA export from the nucleus to
                     the cytoplasm.</body></html>"
                     /label="RRE"
     promoter        7247..7487
                     /note="<html><body>RNA polymerase III promoter for human
                     U6 snRNA</body></html>"
                     /label="U6 promoter"
     misc_feature    7591..7708
                     /note="<html><body>central polypurine tract and central
                     termination sequence of HIV-1</body></html>"
                     /label="cPPT/CTS"
     promoter        7757..507
                     /note="<html><body>human phosphoglycerate kinase 1
                     promoter</body></html>"
                     /label="hPGK promoter"
ORIGIN      
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatctaccat gccagagcca gcgaagtctg
      541 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
      601 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
      661 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaac tcgtttgtga
      721 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct
      781 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
      841 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaaggatc
      901 caccggtcgc caccgcggaa ggatccgtcg ccaggcagcc tgacctcttg acctgcgacg
      961 atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg
     1021 ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg
     1081 gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga
     1141 cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga
     1201 tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg
     1261 agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca
     1321 gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc
     1381 aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact
     1441 tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc
     1501 actatcctgc ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac
     1561 ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca
     1621 ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg
     1681 agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc
     1741 gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc
     1801 gccaagcctg cgagctagtc gcccaggttc tggcctggca gatcggcgtg atggaagagt
     1861 aagtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     1921 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
     1981 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
     2041 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
     2101 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
     2161 ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa
     2221 tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2281 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2341 aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa ctccgcccat
     2401 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     2461 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     2521 cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     2581 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2641 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2701 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     2761 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2821 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2881 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2941 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3001 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3061 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3121 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3181 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3241 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3301 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3361 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3421 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3481 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3541 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3601 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3661 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3721 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3781 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3841 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     3901 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3961 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4021 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4081 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4141 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4201 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4261 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4501 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5041 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5101 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5161 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5221 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5281 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5341 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     5401 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     5461 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     5521 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     5581 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     5641 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     5701 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     5761 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     5821 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     5881 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     5941 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     6001 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     6061 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     6121 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     6181 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     6241 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     6301 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     6361 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     6421 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     6481 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     6541 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     6601 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     6661 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     6721 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     6781 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     6841 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     6901 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     6961 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     7021 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     7081 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     7141 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     7201 acggtatcga tcacgagact agcctcgagc ggccgccccc ttcaccgagg gcctatttcc
     7261 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa
     7321 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     7381 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     7441 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggcaa
     7501 caagatgaag agcaccaact cgagttggtg ctcttcatct tgttgttttt aattctcgac
     7561 ctcgagacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta
     7621 cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca
     7681 aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact
     7741 ttggccgcgg ctcgaggggg
//
