LOCUS       Exported                5130 bp ds-DNA     circular SYN 29-JAN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAK186_2xCBX7(1-176)-IFP2.0C
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5130)
  AUTHORS   Jeltsch Group
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 29, 2024 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5130
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    525..612
                     /label=sequenced
     misc_feature    613..840
                     /label=CBX7CD
     misc_feature    706..708
                     /label=W32A
     misc_feature    715..717
                     /label=W35A
     misc_feature    841..849
                     /label=18aa C50 linker
                     /label=50aa C50 linker
                     /note="Copied from rg2104_ChePep3_hRas"
     misc_feature    841..849
                     /label=sequenced
     CDS             850..870
                     /codon_start=1
                     /label=NLS
                     /translation="PKKKRKV"
     misc_feature    867..876
                     /label=sequenced
     misc_feature    877..1104
                     /label=CBX7CD
     misc_feature    970..972
                     /label=W32A
     misc_feature    979..981
                     /label=W35A
     misc_feature    1105..1158
                     /label=18aa C50 linker
                     /label=50aa C50 linker
                     /note="Copied from rg2104_ChePep3_hRas"
     misc_feature    1105..1158
                     /label=sequenced
     CDS             1158..1725
                     /codon_start=1
                     /product="bacteriophytochrome-based monomeric infrared 
                     fluorescent protein (Yu et al., 2014)"
                     /label=IFP2.0
                     /translation="DSIGPHALRNAMFALESAPNLRALAEVATQTVRELSGFDRVMLYK
                     FAPDATGEVIAEARREGMQAYLGHRFPASTTPAQARALYTRHLLRLTADTRAAAVPLDP
                     VLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGGQLWGLIVCHHQTPY
                     VLPPDLRTTLEYLGRLLSLQVQRKEA"
     misc_feature    1159..1725
                     /label=C133
     misc_feature    1726..1728
                     /label=Synthetic stop
     misc_feature    1729..1794
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    1803..1813
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     polyA_signal    1918..2039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2046..2501)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2528..2632
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2634..2991
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2842..2977
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3026..3820
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4052..4099
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    4052..4099
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      4428..5016
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggagct gtcagccatc ggcgagcagg tgttcgccgt ggagagcatc
      661 cggaagaagc gcgtgcggaa gggtaaagtc gagtatctgg tgaagtggaa aggatggccc
      721 ccaaagtaca gcacgtggga gccagaagag cacatcttgg acccccgcct cgtcatggcc
      781 tacgaggaga aggaggagag agaccgagca tcggggtata ggaagagagg tccgaaaccc
      841 actggatcgc ccaagaaaaa acgcaaggtg gccaccatgg agctgtcagc catcggcgag
      901 caggtgttcg ccgtggagag catccggaag aagcgcgtgc ggaagggtaa agtcgagtat
      961 ctggtgaagt ggaaaggatg gcccccaaag tacagcacgt gggagccaga agagcacatc
     1021 ttggaccccc gcctcgtcat ggcctacgag gagaaggagg agagagaccg agcatcgggg
     1081 tataggaaga gaggtccgaa acccactgga tcgtcacaaa cctcgggttc atccacgtcg
     1141 gaaggccagg agggcacgga ctctattgga ccccacgctc tgagaaatgc aatgttcgct
     1201 ctggagagtg ctcctaatct ccgggcactg gctgaagtcg caacccaaac agtccgggaa
     1261 ctgtcaggtt tcgaccgggt gatgctgtac aagtttgcac cagacgcaac aggagaggtt
     1321 attgccgaag caaggcgcga gggcatgcag gcttacctcg ggcataggtt tcccgcatcc
     1381 accacccctg cacaagctag ggccctctac acaagacacc tgctccggct gaccgcagac
     1441 accagggctg cagcagtgcc cctcgacccc gtgctgaatc cccagacaaa tgctcctaca
     1501 cctctgggcg gagctgtcct cagagctaca tccccaatgc acatgcagta cctgaggaat
     1561 atgggagtgg gctcctccct gagcgtcagc gtcgtggtcg gcggccagct gtggggactg
     1621 attgtctgcc accatcagac accctacgtg ctgccacccg atctgcggac caccctggag
     1681 tatctgggga ggctcctgtc cctgcaggtg cagaggaaag aagcctaatc cggactcaga
     1741 tctcgagctc aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga
     1801 tctagataac tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     1861 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     1921 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     1981 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2041 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2101 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2161 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2221 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2281 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2341 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2401 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2461 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     2521 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2581 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     2641 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2701 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2761 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2821 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2881 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2941 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3001 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3061 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3121 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3181 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     3241 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3301 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3361 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3421 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3481 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3541 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3601 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3661 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3721 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3781 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3841 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     3901 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     3961 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4021 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4081 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     4141 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4201 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4261 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4321 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     4381 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4441 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4501 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4561 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4621 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4681 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4741 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4801 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4861 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4921 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4981 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5041 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     5101 tctgtggata accgtattac cgccatgcat
//
