LOCUS       pRC1062                 7217 bp ds-DNA     circular     16-AUG-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     misc_feature    1..8
                     /label="SbfI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    9..342
                     /label="CMV enhancer (human)"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     promoter        349..1709
                     /label="Actin promoter (human)"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          1667..1688
                     /label="oRC0262.pCAG-F_seq F"
                     /note="sequence: GGGCAACGTGCTGGTTATTGTG"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    1710..1717
                     /label="NotI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     promoter        1718..1750
                     /label="T7 promoter"
                     /ApEinfo_revcolor=#84b0dc
                     /ApEinfo_fwdcolor=#84b0dc
     misc_feature    1751..1756
                     /label="HpaI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    1757..1781
                     /label="polyA25 spacer"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    1782..1801
                     /label="Outer HomologyArm"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     intron          1808..2063
                     /label="3' Intron"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     primer          1958..1980
                     /label="oRC0718.3'intron-retained_seq F"
                     /note="sequence: GGAGATATAGTCTGCTCTGCATG"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    2040..2063
                     /label="3' intron retained"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     misc_feature    2064..2083
                     /label="Inner HomologyArm"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    2084..2133
                     /label="PABPspacer50v3"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    2123..2133
                     /label="3-Frame stop"
                     /ApEinfo_revcolor=#d59687
                     /ApEinfo_fwdcolor=#d59687
     IRES            2140..2805
                     /label="synIRES-RC25 (iHRVB3-based)"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2531..2570
                     /label="Apt17 (eIF4G binding)"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     primer          2714..2735
                     /label="oRC1651.iHRV-B3_seq F"
                     /note="sequence: GGGACGGGACCGACTACTTTGG"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     CDS             2806..3525
                     /label="Translation 2806-3525"
     misc_feature    2815..3519
                     /label="mNeonGreen.v2"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     misc_feature    3526..3531
                     /label="MfeI"
                     /ApEinfo_revcolor=#b1ff67
                     /ApEinfo_fwdcolor=#b1ff67
     misc_feature    3532..3632
                     /label="Moderna 3'UTRv2  (M. Richner et al., Cell, 2017)"
                     /ApEinfo_revcolor=#d6b295
                     /ApEinfo_fwdcolor=#d6b295
     misc_feature    complement(3633..3652)
                     /label="Inner HomologyArm RevComp"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    3653..3674
                     /label="DS intron retained"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
     intron          3653..3859
                     /label="T4.td DS Intron"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     primer          complement(3735..3756)
                     /label="oRC0719.5'intron-retained_seq R"
                     /note="sequence: TTAGCACGGGATTGTCTACTAG"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    complement(3866..3885)
                     /label="Outer HomologyArm RevComp"
                     /ApEinfo_revcolor=#c7b0e3
                     /ApEinfo_fwdcolor=#c7b0e3
     misc_feature    3886..3910
                     /label="polyA25 spacer"
                     /ApEinfo_revcolor=#f8d3a9
                     /ApEinfo_fwdcolor=#f8d3a9
     misc_feature    3911..3918
                     /label="NotI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     terminator      3919..3966
                     /label="T7 terminator"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     misc_feature    3967..3972
                     /label="NheI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     misc_feature    3973..3980
                     /label="FseI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     polyA_signal    3981..4205
                     /label="BGH pA"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     primer          complement(4041..4060)
                     /label="oRC1571.BGHpA_seq R"
                     /note="sequence: CAGTGGGAGTGGCACCTTCC"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    4206..4213
                     /label="AscI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     terminator      complement(4214..4586)
                     /label="SV40 p(A) terminator"
                     /ApEinfo_revcolor=#ff9ccd
                     /ApEinfo_fwdcolor=#ff9ccd
     CDS             complement(4620..5216)
                     /label="PuroR"
                     /ApEinfo_revcolor=#faac61
                     /ApEinfo_fwdcolor=#faac61
     misc_feature    complement(5217..5555)
                     /label="SV40 promoter-enhancer-origin"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     ori             complement(5241..5376)
                     /label="SV40 origin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     promoter        complement(5275..5543)
                     /label="promoter"
                     /ApEinfo_revcolor=#85dae9
                     /ApEinfo_fwdcolor=#85dae9
     misc_feature    complement(5340..5555)
                     /label="SV40 enhancer"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    5556..5561
                     /label="SacI"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
     terminator      complement(5562..5588)
                     /label="KanR Terminator"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     CDS             complement(5589..6404)
                     /label="KanR"
                     /ApEinfo_revcolor=#75c6a9
                     /ApEinfo_fwdcolor=#75c6a9
     promoter        complement(6405..6552)
                     /label="KanR Promoter"
                     /ApEinfo_revcolor=#82ca9d
                     /ApEinfo_fwdcolor=#82ca9d
     rep_origin      6629..7217
                     /label="ColE1"
                     /ApEinfo_revcolor=#9eafd2
                     /ApEinfo_fwdcolor=#9eafd2
ORIGIN
        1 cctgcaggTT ATTAATAGTA ATCAATTAcg gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc
      421 ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg
      481 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
      541 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
      601 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      661 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      721 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      781 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      841 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      901 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      961 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     1021 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     1081 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     1141 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     1201 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     1261 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     1321 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     1381 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     1441 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     1501 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1561 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1621 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1681 ttattgtgct gTCTCATCAT TTTGGCAAAG Cggccgcggc cagtgaattg taatacGACT
     1741 CACTATAGGG GTTAACAAAA AAAAAAAAAA AAAAAAAAAA ATCACCGACC GATCTATCTG
     1801 AACCGGTggg aattctagag aaaatttcgt ctggattagt tacttatcgt gtaaaatctg
     1861 ataaatggaa ttggttctac ataaatgcct aacgactatc cctttgggga gtagggtcaa
     1921 gtgactcgaa acgatagaca acttgcttta acaagttgga gatatagtct gctCTGCATG
     1981 GTGACATGCA Gctggatata attccggggt aagattaacg accttatctg aacataacgc
     2041 taccgtttaa tattgcgtca tattcggcga ccatttgtgt ggtaaaaaaa aaaaaccaaa
     2101 aaaaaaaaac aaaaaaaaaa aataattgac taagaTATCt taaaacagcg gatgggtacc
     2161 ccaccatccg acccactggg tgtagtactc tggtacttcg tacctttgta cgcctgttct
     2221 tcccattgta cccttcctga acttccaacc caagtaacgt tagaagctca acatttagta
     2281 caacaggaag caccacatcc agtggtgttt agtacaagca cttctgtttc cccggagcga
     2341 ggtataggct gtacccactg ccaaaaacct ttaaccgtta tccgccaacc aactacgtaa
     2401 aagctagtag tattatgttt ttaactaggc gttcgatcag gtggatttcc cctccactag
     2461 tttggtcgat gaggctagga attccccacg ggtgaccgtg tcctagcctg cgtggcggcc
     2521 aacccagcCC ACTCACTATT TGTTTTCGCG CCCAGTTGCA AAAAGTGTCG GGgctgggac
     2581 gcctttttat agacatggtg tgaagactcg catgtgcttg gttgtgattc ctccggcccc
     2641 tgaatgcggc taaccttaac cctggagcct tgtgtcacaa accagtgatg ataaggtcgt
     2701 aatgagcaat tccgggacgg gaccgactac tttgggtgtc cgtgtttctt atttttctta
     2761 ttattgtctt atggtcacag catatatata acatatactg tgatcATGGG ATCCgtgtct
     2821 aaaggagagg aagataacat ggcgagcctc ccagccactc atgaactcca catattcgga
     2881 agcatcaacg gggtggactt tgacatggta ggtcagggga ccggcaatcc caacgacggt
     2941 tacgaggaat tgaatctcaa gtccactaag ggcgacctcc agttttctcc gtggattctc
     3001 gtcccacata tcgggtatgg tttccaccaa tacttgcctt acccggatgg aatgtcaccg
     3061 ttccaggctg ccatggttga tggctctggt tatcaggtac acagaacgat gcaatttgag
     3121 gacggagcgt cattgactgt aaactacaga tacacatacg aaggctctca cataaagggc
     3181 gaagcacaag ttaaaggcac gggattcccc gctgatggtc ctgtcatgac aaattctctt
     3241 accgcagccg actggtgtcg atccaagaaa acgtatccca atgacaaaac tatcatatct
     3301 acgttcaaat ggtcatatac aacgggaaac ggtaagagat accggagcac tgccagaacc
     3361 acctatacat ttgccaaacc aatggcggct aactacctga agaaccagcc catgtatgtg
     3421 ttcaggaaaa cggaactgaa acactccaag acagagctga atttcaagga atggcaaaaa
     3481 gccttcacgg atgttatggg gatggatgag ctgtataaaT AGTaacaatt ggctggagcc
     3541 tcggtggcca tgcttcttgc cccttgggcc tccccccagc ccctcctccc cttcctgcac
     3601 ccgtaccccc gtggtctttg aataaagtct gaaccacaca aatggtcgcc gactcagtag
     3661 atgttttctt gggttaattg aggcctgagt ataaggtgac ttatacttgt aatctatcta
     3721 aacggggaac ctctctagta gacaatcccg tgctaaattg taggactgcc ctttaataaa
     3781 tacttctata tttaaagagg tatttatgaa aagcggaatt tatcagatta aaaatacttt
     3841 ctctagagtc gacctgcagA CGCGTTCAGA TAGATCGGTC GGTGAAAAAA AAAAAAAAAA
     3901 AAAAAAAAAA GCGGCCGCCT AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT
     3961 TTTTTGGCTA GCGGCCGGCC TGTGCCTTCT AGTTGCCAGC catctgttgt ttgcccctcc
     4021 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     4081 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     4141 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     4201 atggcggcgc gccactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4261 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     4321 ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4381 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4441 gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     4501 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     4561 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaatcag
     4621 gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg
     4681 gtgacggtga agccgagccg ctcgtagaag gggaggttgc ggggcgcgga ggtTTCcagg
     4741 aaggcgggca ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc
     4801 agacccttgc cctggtggtc gggcgaCACg ccgacggtgg ccaggaacca cgcgggctcc
     4861 ttgggccggt gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg
     4921 ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc
     4981 ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg
     5041 agcccgacgc gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc
     5101 ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg
     5161 gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatgcgg
     5221 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
     5281 agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
     5341 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
     5401 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     5461 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     5521 ggagcctggg gACTTTCCAC ACCCTAACTG ACACAGAGCT Cgtgttacaa ccaattaacc
     5581 aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt catatcagga
     5641 ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg
     5701 cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg tccaacatca
     5761 atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa atcaccatga
     5821 gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca gacttgttca
     5881 acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc gttattcatt
     5941 cgtgattgcg cctgagcgag gcgaaatacg cgatcgctgt taaaaggaca attacaaaca
     6001 ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt ttcacctgaa
     6061 tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac
     6121 catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat aaattccgtc
     6181 agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc tttgccatgt
     6241 ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt cgcacctgat
     6301 tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt
     6361 aatcgcggcc tggagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     6421 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     6481 taacatcaga gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag
     6541 ttgaaggatc agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     6601 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6661 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6721 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6781 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6841 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6901 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6961 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7021 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7081 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7141 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7201 ggcggagcct atggaaa
//