LOCUS       Exported                4574 bp ds-DNA     circular SYN 29-MAR-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pA-RFP-rA1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4574)
  AUTHORS   Biochemicals, Synthetic Biology Research Center
  TITLE     Direct Submission
  JOURNAL   Exported Mar 29, 2022 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4574
                     /organism="PCR product"
                     /mol_type="other DNA"
                     /label=/vntifkey=98
                     /note="/vntifkey=98"
                     /note="color: #ffffff"
     source          3953..3972
                     /organism="PCR product"
                     /mol_type="other DNA"
                     /note="/vntifkey=98"
                     /note="color: #ffffff"
     CDS             complement(885..1544)
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="CmR confers resistance to chloramphenicol"
                     /note="/vntifkey=4"
                     /note="color: #a6acb3; direction: LEFT"
     rep_origin      complement(2173..2717)
                     /direction=LEFT
                     /label=p15a origin
                     /label=Rep_Origin_1
                     /note="p15A ori Plasmids containing the medium-copy-number 
                     p15A origin of replication can be propagated in E. coli 
                     cells that contain a second plasmid with the ColE1 origin."
                     /note="/vntifkey=33"
                     /note="color: #ffff00"
     misc_feature    2893..2922
                     /label=Ptrc
                     /label=Misc_Feature_3
                     /note="trc promoter strong E. coli promoter; hybrid between
                     the trp and lac UV5 promoters"
                     /note="/vntifkey=21"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2928..2948
                     /label=lacO
                     /note="color: #ffffff"
     CDS             2995..3705
                     /product="monomeric derivative of DsRed fluorescent 
                     protein"
                     /label=RFP
                     /label=CDS_1
                     /note="mCherry mammalian codon-optimized"
                     /note="/vntifkey=4"
                     /note="color: #ff0000; direction: RIGHT"
     primer_bind     complement(3710..3736)
                     /label=mCherry-VF
                     /note="color: black; sequence: tgcatgcctgcaggtcgactctagagg"
     primer_bind     3717..3756
                     /label=sgRNA(RFP)-F
                     /note="color: black; sequence: 
                     agtcgacctgcaggcatgcataaggatctccaggcatcaa"
     promoter        3918..3952
                     /label=PJ23119
                     /label=J23100
                     /note="/vntifkey=30"
                     /note="color: #ffffff"
     misc_feature    3953..4054
                     /label=sgRNA
                     /note="color: #a6acb3"
     misc_feature    3953..3972
                     /label=Spacer
                     /note="color: #ffff99"
     primer_bind     complement(4044..4083)
                     /label=sgRNA(RFP)-R
                     /note="color: black; sequence: 
                     ctcatccgccaaaacagccacccgggtcaaaaaaagcacc"
     primer_bind     4064..4088
                     /label=mCherry-VR
                     /note="color: black; sequence: tggctgttttggcggatgagagaag"
ORIGIN
        1 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
       61 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
      121 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
      181 ggtttccgtg tttcgtaaag tctggaaacg cggaagtccc ctacgtgctg ctgaagttgc
      241 ccgcaacaga gagtggaacc aaccggtgat accacgatac tatgactgag agtcaacgcc
      301 atgagcggcc tcatttctta ttctgagtta caacagtccg caccgctgtc cggtagctcc
      361 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
      421 caactcgtag gacaggtgcc ggcagcgccc aacagtcccc cggccacggg gcctgccacc
      481 atacccacgc cgaaacaagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
      541 tgctggctac cctgtggaac acctacatct gtattaacga agcgctaacc gtttttatca
      601 ggctctggga ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct
      661 gagcaaactg gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata
      721 aaccggtaaa ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga
      781 ccgggtcgaa tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc
      841 gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
      901 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacag
      961 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     1021 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     1081 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1141 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1201 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1261 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1321 atacggaact ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     1381 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     1441 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     1501 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     1561 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     1621 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     1681 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     1741 ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg
     1801 atggtgtttt tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc
     1861 tactgacggg gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt
     1921 atactggctt actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga
     1981 gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct
     2041 cgctcactga ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac
     2101 ggggcggaga tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc
     2161 ggcaaagccg tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc
     2221 aaatcagtgg tggcgaaacc cgacaggact ataaagatac caggcgtttc ccctggcggc
     2281 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
     2341 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     2401 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     2521 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     2581 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     2641 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     2701 gaccaaaacg atctcaagaa gatcatctta ttaatcagat aaaatatttc aagatttcag
     2761 tgcaatttat ctcttcaaat gtagcacctg aagtcagccc taattcgtgt cgctcaaggc
     2821 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
     2881 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
     2941 taacaatttc gaattcaaaa gatctatcga tcgaggatcc aggaggtaca atcaatggtg
     3001 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     3061 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     3121 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     3181 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     3241 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     3301 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     3361 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     3421 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     3481 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     3541 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     3601 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     3661 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaggtac ctctagagtc
     3721 gacctgcagg catgcataag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     3781 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     3841 ggctcacctt cgggtgggcc tttctgcgtt tatacctagg gatatattcc gcttcctcgg
     3901 cgaccggtta aagatctttg acagctagct cagtcctagg tataatacta gtagacgctg
     3961 attacatgga ctgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
     4021 acttgaaaaa gtggcaccga gtcggtgctt tttttgaccc gggtggctgt tttggcggat
     4081 gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt ctgataaaac
     4141 agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg aactcagaag
     4201 tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     4261 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     4321 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     4381 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     4441 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactcg gatcccatgg
     4501 tacgcgttga ttctccttac gcatctgtgc ggtatttcac accgcatgta ctagttaact
     4561 tcaggacctg cagc
//
