LOCUS       pPBpuro-miRFP670-eDHFR( 9682 bp ds-DNA     circular     07-NOV-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     rep_origin      complement(135..441)
                     /label="F1 ori"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(460..528)
                     /label="LacZ alpha"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     primer_bind     599..616
                     /label="M13-fwd"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer_bind     625..645
                     /label="T7"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    1009..2726
                     /label="CAG promoter"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             2756..5515
                     /label="Translation 2756-5515"
     misc_feature    2759..3700
                     /label="miRFP670"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    3740..4234
                     /label="eDHFR(69K6)"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     CDS             4244..4375
                     /label="Linker(44aa)"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     CDS             4376..5512
                     /label="Gs(alpha)_C3S"
                     /ApEinfo_revcolor=#b7e6d7
                     /ApEinfo_fwdcolor=#b7e6d7
     misc_feature    4382..4384
                     /label="C->S mutation"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     promoter        5587..6133
                     /label="IRES"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     CDS             6143..6736
                     /label="puromycin N-acetyltransferase"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     CDS             6143..6736
                     /label="Translation 6143-6736"
     primer_bind     complement(7039..7059)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_binding    complement(7065..7087)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(7091..7121)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     primer_bind     complement(7492..7511)
                     /label="T3"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     promoter        complement(7494..7512)
                     /label="T3 promoter"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     primer_bind     complement(7529..7549)
                     /label="M13-rev"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_binding    complement(7555..7577)
                     /label="LacO"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        complement(7581..7611)
                     /label="lac promoter"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     rep_origin      complement(7917..8599)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     CDS             complement(8697..9356)
                     /label="AmpR"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat
      721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg
      781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact
      841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta
      901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat
      961 atattttctt gttatagata tcaactagaa tgctagcatg ggcccatctc gacattgatt
     1021 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     1081 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     1141 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     1201 acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     1261 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     1321 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     1381 tattaccatg ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     1441 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     1501 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     1561 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     1621 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     1681 ttgccttcgc cccgtgcccc gctccgcgcc gcctcgcgcc gcccgccccg gctctgactg
     1741 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     1801 cgcttggttt aatgacggct cgtttctttt ctgtggctgc gtgaaagcct taaagggctc
     1861 cgggagggcc ctttgtgcgg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
     1921 ggggagcgcc gcgtgcggcc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     1981 gggctttgtg cgctccgcgt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     2041 gcgggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     2101 agggggtgtg ggcgcggcgg tcgggctgta acccccccct gcacccccct ccccgagttg
     2161 ctgagcacgg cccggcttcg ggtgcggggc tccgtgcggg gcgtggcgcg gggctcgccg
     2221 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     2281 gggagggctc gggggagggg cgcggcggcc ccggagcgcc ggcggctgtc gaggcgcggc
     2341 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     2401 ccaaatctgg cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg
     2461 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     2521 gccgccgtcc ccttctccat ctccagcctc ggggctgccg cagggggacg gctgccttcg
     2581 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     2641 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttgt
     2701 tgtgctgtct catcattttg gcaaagaatt GGGATCCTCA CGTCAATTGG CCACCatggt
     2761 agcaggtcat gcctctggca gccccgcatt cgggaccgcc tctcattcga attgcgaaca
     2821 tgaagagatc cacctcgccg gctcgatcca gccgcatggc gcgcttctgg tcgtcagcga
     2881 acatgatcat cgcgtcatcc aggccagcgc caacgccgcg gaatttctga atctcggaag
     2941 cgtactcggc gttccgctcg ccgagatcga cggcgatctg ttgatcaaga tcctgccgca
     3001 tctcgatccc accgccgaag gcatgccggt cgcggtgcgc tgccggatcg gcaatccctc
     3061 tacggagtac tgcggtctga tgcatcggcc tccggaaggc gggctgatca tcgaactcga
     3121 acgtgccggc ccgtcgatcg atctgtcagg cacgctggcg ccggcgctgg agcggatccg
     3181 cacggcgggt tcactgcgcg cgctgtgcga tgacaccgtg ctgctgtttc agcagtgcac
     3241 cggctacgac cgggtgatgg tgtatcgttt cgatgagcaa ggccacggcc tggtattctc
     3301 cgagtgccat gtgcctgggc tcgaatccta tttcggcaac cgctatccgt cgtcgactgt
     3361 cccgcagatg gcgcggcagc tgtacgtgcg gcagcgcgtc cgcgtgctgg tcgacgtcac
     3421 ctatcagccg gtgccgctgg agccgcggct gtcgccgctg accgggcgcg atctcgacat
     3481 gtcgggctgc ttcctgcgct cgatgtcgcc gtgccatctg cagttcctga aggacatggg
     3541 cgtgcgcgcc accctggcgg tgtcgctggt ggtcggcggc aagctgtggg gcctggttgt
     3601 ctgtcaccat tatctgccgc gcttcatccg tttcgagctg cgggcgatct gcaaacggct
     3661 cgccgaaagg atcgcgacgc ggatcaccgc gcttgagagc CTcTACAAGT CTGGGTCCGC
     3721 AGCCAGTAGC GCCGGAAGTA TGATCAGTCT GATTGCGGCG TTAGCGGTAG ATCGCGTTAT
     3781 CGGCATGGAA AACGCCATGC CGTGGAACCT GCCTGCCGAT CTCGCCTGGT TTAAACGCAA
     3841 CACCTTAAAT AAACCCGTGA TTATGGGCCG CCATACCTGG GAATCAATCG GTCGTCCGTT
     3901 GCCAGGACGC AAAAATATTA TCCTCAGCAG TCAACCGGGT ACGGACAAAA AAAAGAAAAA
     3961 GAAAGATCGC GTAACGTGGG TGAAGTCGGT GGATGAAGCC ATCGCGGCGT GTGGTGACGT
     4021 ACCAGAAATC ATGGTGATTG GCGGCGGTCG CGTTTATGAA CAGTTCTTGC CAAAAGCGCA
     4081 AAAACTGTAT CTGACGCATA TCGACGCAGA AGTGGAAGGC GACACCCATT TCCCGGATTA
     4141 CGAGCCGGAT GACTGGGAAT CGGTATTCAG CGAATTCCAC GATGCTGATG CGCAGAACTC
     4201 TCACAGCTAT TGCTTTGAGA TTCTGGAGCG GCGGCTGTAC AAGTCCGGAC TCAGATCTCG
     4261 ACAAGGTAGT GGTGCTGGCT CTGGTGCTGG TAGTGGCGCT GGTTCCGGTG CTGGCTCTGG
     4321 CGCGCCTCGA GCTCAAGCTT CGAATTCTGC AGTCGACGGT ACCGCGGGCC CGGGtATGGG
     4381 CaGCCTCGGC AACAGTAAGA CCGAGGACCA GCGCAACGAG GAGAAGGCGC AGCGCGAGGC
     4441 CAACAAAAAG ATCGAGAAGC AGCTGCAGAA GGACAAGCAG GTCTACCGGG CCACGCACCG
     4501 CCTGCTGCTG CTGGGTGCTG GAGAGTCTGG CAAAAGCACC ATTGTGAAGC AGATGAGGAT
     4561 CCTACATGTT AATGGGTTTA ACGGAGACGA GAAGGCCACC AAAGTGCAGG ACATCAAAAA
     4621 CAACCTGAAG GAGGCCATTG AAACCATTGT GGCCGCCATG AGCAACCTGG TGCCCCCCGT
     4681 GGAGCTGGCC AACCCTGAGA ACCAGTTCAG AGTGGACTAC ATTCTGAGCG TGATGAACGT
     4741 GCCAAACTTT GACTTCCCAC CTGAATTCTA TGAGCATGCC AAGGCTCTGT GGGAGGATGA
     4801 GGGAGTTCGT GCCTGCTACG AGCGCTCCAA CGAGTACCAG CTGATCGACT GTGCCCAGTA
     4861 CTTCCTGGAC AAGATTGATG TGATCAAGCA GGCCGACTAC GTGCCAAGTG ACCAGGACCT
     4921 GCTTCGCTGC cGcGTCCTGA CCTCTGGAAT CTTTGAGACC AAGTTCCAGG TGGACAAAGT
     4981 CAACTTCCAC ATGTTCGATG TGGGCGGCCA GCGCGATGAA CGCCGCAAGT GGATCCAGTG
     5041 CTTCAATGAT GTGACTGCCA TCATCTTCGT GGTGGCCAGC AGCAGCTACA ACATGGTCAT
     5101 CCGGGAGGAC AACCAGACCA ACCGTCTGCA GGAGGCTCTG AACCTCTTCA AGAGCATCTG
     5161 GAACAACAGA TGGCTGCGTA CCATCTCTGT GATCCTCTTC CTCAACAAGC AAGATCTGCT
     5221 TGCTGAGAAG GTCCTCGCTG GGAAATCGAA GATTGAGGAC TACTTTCCAG AGTTCGCTCG
     5281 CTACACCACT CCTGAGGATG CGACTCCCGA GCCCGGAGAG GACCCACGCG TGACCCGGGC
     5341 CAAGTACTTC ATCCGGGATG AGTTTCTGAG AATCAGCACT GCTAGTGGAG ATGGACGTCA
     5401 CTACTGCTAC CCTCACTTTA CCTGCGCCGT GGACACTGAG AACATCCGCC GTGTCTTCAA
     5461 CGACTGCCGT GACATCATCC AGCGCATGCA TCTTCGCCAA TACGAGCTGC TCTAAACCGG
     5521 GATAAgtcAA CTAACTTAAG CTAGCAACGG TTTCCCTCTA GCGGGATCAA TTCCGCCCCC
     5581 CCCCCCTAAC GTTACTGGCC GAAGCCGCTT GGAATAAGGC CGGTGTGCGT TTGTCTATAT
     5641 GTTATTTTCC ACCATATTGC CGTCTTTTGG CAATGTGAGG GCCCGGAAAC CTGGCCCTGT
     5701 CTTCTTGACG AGCATTCCTA GGGGTCTTTC CCCTCTCGCC AAAGGAATGC AAGGTCTGTT
     5761 GAATGTCGTG AAGGAAGCAG TTCCTCTGGA AGCTTCTTGA AGACAAACAA CGTCTGTAGC
     5821 GACCCTTTGC AGGCAGCGGA ACCCCCCACC TGGCGACAGG TGCCTCTGCG GCCAAAAGCC
     5881 ACGTGTATAA GATACACCTG CAAAGGCGGC ACAACCCCAG TGCCACGTTG TGAGTTGGAT
     5941 AGTTGTGGAA AGAGTCAAAT GGCTCTCCTC AAGCGTATTC AACAAGGGGC TGAAGGATGC
     6001 CCAGAAGGTA CCCCATTGTA TGGGATCTGA TCTGGGGCCT CGGTGCACAT GCTTTACATG
     6061 TGTTTAGTCG AGGTTAAAAA ACGTCTAGGC CCCCCGAACC ACGGGGACGT GGTTTTCCTT
     6121 TGAAAAACAC GATAATACCA TGACCGAGTA CAAGCCCACG GTGCGCCTCG CCACCCGCGA
     6181 CGACGTCCCC aGGGCCGTAC GCACCCTCGC CGCCGCGTTC GCCGACTACC CCGCCACGCG
     6241 CCACACCGTC GAtCCGGACC GCCACATCGA GCGGGTCACC GAGCTGCAAG AACTCTTCCT
     6301 CACGCGCGTC GGGCTCGACA TCGGCAAGGT GTGGGTCGCG GACGACGGCG CCGCGGTGGC
     6361 GGTCTGGACC ACGCCGGAGA GCGTCGAAGC GGGGGCGGTG TTCGCCGAGA TCGGCCCGCG
     6421 CATGGCCGAG TTGAGCGGTT CCCGGCTGGC CGCGCAGCAA CAGATGGAAG GCCTCCTGGC
     6481 GCCGCACCGG CCCAAGGAGC CCGCGTGGTT CCTGGCCACC GTCGGCGTCT CGCCCGACCA
     6541 CCAGGGCAAG GGTCTGGGCA GCGCCGTCGT GCTCCCCGGA GTGGAGGCGG CCGAGCGCGC
     6601 CGGGGTGCCC GCCTTCCTGG AGACCTCCGC GCCCCGCAAC CTCCCCTTCT ACGAGCGGCT
     6661 CGGCTTCACC GTCACCGCCG ACGTCGAGTG CCCGAAGGAC CGCGCGACCT GGTGCATGAC
     6721 CCGCAAGCCC GGTGCCTGAT cgatgatcta gagctcgctg atcagcctcg actgtgcctt
     6781 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     6841 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     6901 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     6961 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaacctgca
     7021 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7081 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7141 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7201 cgtgccagcg gatctTAAAA GTTTTGTTAC TTTATAGAAG AAATTTTGAG TTTTTGTTTT
     7261 TTTTTAATAA ATAAATAAAC ATAAATAAAT TGTTTGTTGA ATTTATTATT AGTATGTAAG
     7321 TGTAAATATA ATAAAACTTA ATATCTATTC AAATTAATAA ATAAACCTCG ATATACAGAC
     7381 CGATAAAACA CATGCGTCAA TTTTACGCAT GATTATCTTT AACGTACGTC ACAATATGAT
     7441 TATCTTTCTA GGGTTAATCT AGTATACGCG Tttaattaat ccagcttttg ttccctttag
     7501 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     7561 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7621 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7681 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7741 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7801 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7861 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7921 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7981 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     8041 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     8101 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     8161 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     8221 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8281 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8341 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     8401 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8461 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     8521 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     8581 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     8641 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     8701 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     8761 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     8821 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8881 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8941 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     9001 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     9061 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     9121 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     9181 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     9241 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     9301 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     9361 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     9421 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     9481 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     9541 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     9601 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     9661 acatttcccc gaaaagtgcc ac
//