LOCUS       sequence-252820-        6121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of frp in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6121)
  AUTHORS   Gregor C, Pape JK, Gwosch KC, Gilat T, Sahl SJ, Hell SW
  TITLE     Autonomous bioluminescence imaging of single mammalian cells with 
            the bacterial bioluminescence system.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Dec 2. pii: 1913616116. doi: 
            10.1073/pnas.1913616116.
  PUBMED    31792180
REFERENCE   2  (bases 1 to 6121)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2019 Dec 2. pii: 1913616116. doi: 
            10.1073/pnas.1913616116."
COMMENT     Sequence Label: co frp pcDNA3.1(+)
FEATURES             Location/Qualifiers
     source          1..6121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1712..1729)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1718..1942
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1988..2416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2075..2094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2285..2306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2425..2445)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2430..2759
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2610..2745
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2672..2691
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2826..3620
                     /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(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2880..2899)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3490..3509
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3794..3915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3831..3850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3885..3904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3964..3980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3977..3999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3988..4004
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4012..4018,4019..4036,4037..4042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4057..4078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4195..4212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4366..4954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4446..4465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5125..5916,5917..5985))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5748..5767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5986..6090)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 atggtgaaga tccagcccat ccccaccacc agccagggca gcctgttcat catgaacagc
      961 accatcgaga ccatcctggg ccaccgcagc atccgcaagt tcaccagcga gcccatcgcc
     1021 agcgagcagc tgcagaccat cctgcagagc ggcctggccg ccagcagcag cagcatgctg
     1081 caggtggtga gcatcatccg cgtgaccgac accgagaagc gcaagctgct ggcccagtac
     1141 gccggcaacc agacctacgt ggagagcgcc gccgagttcc tggtgttctg catcgactac
     1201 cagcgccacg ccaccatcaa ccccgacgtg caggccgact tcaccgagct gaccctgatc
     1261 ggcgccgtgg acagcggcat catggcccag aactgcctgc tggccgccga gagcatgggc
     1321 ctgggcggcg tgtacatcgg cggcctgcgc aacagcgccg cccaggtgga cgagctgctg
     1381 ggcctgccca agaacaccgc catcctgttc ggcatgtgcc tgggccaccc cgaccagagc
     1441 cccgagacca agccccgcct gcccgcccac gtgatcgtgc acgagaacca gtaccaggcc
     1501 ctgaacatcg acgacgtgca ggcctacgac aagaccatgc aggagtacta cgccagccgc
     1561 accagcaacc agaagcagag cgtgtggagc caggagaccg ccggcaagct ggccggcgag
     1621 agccgccccc acatcctgcc ctacctgaac agcaagggcc tggccaagcg ctgactcgag
     1681 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     1741 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     1801 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     1861 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     1921 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     1981 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2041 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2101 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     2161 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     2221 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     2281 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     2341 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     2401 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     2461 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     2521 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2581 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     2641 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     2701 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     2761 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     2821 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2881 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2941 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     3001 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     3061 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     3121 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     3181 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     3241 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     3301 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     3361 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3421 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3481 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3541 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3601 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3661 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3721 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3781 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3841 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3901 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     3961 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     4021 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4081 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4141 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4201 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4261 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4321 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4381 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4861 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6121 c
//
