LOCUS       sequence-151190-        7247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses smURFP and Heme Oxygenase-1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7247)
  AUTHORS   Rodriguez EA, Tran GN, Gross LA, Crisp JL, Shu X, Lin JY, Tsien RY
  TITLE     A far-red fluorescent protein evolved from a cyanobacterial 
            phycobiliprotein.
  JOURNAL   Nat Methods. 2016 Aug 1. doi: 10.1038/nmeth.3935.
  PUBMED    27479328
REFERENCE   2  (bases 1 to 7247)
  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: "Nat 
            Methods. 2016 Aug 1. doi: 10.1038/nmeth.3935."
COMMENT     Sequence Label: pcDNA3-smURFP-IRES-HO-1
FEATURES             Location/Qualifiers
     source          1..7247
                     /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"
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             923..1324
                     /codon_start=1
                     /product="dimeric small ultra-red fluorescent protein 
                     derived from a cyanobacterial phycobiliprotein (Rodriguez 
                     et al., 2016)"
                     /label=smURFP
                     /translation="MAKTSEQRVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAF
                     SQRDRALCLRDYGWFLHLITFCLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRC
                     LKEASLSLLDEEDANETAPYFDYIIKAMS"
     primer_bind     complement(2672..2689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2678..2902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2948..3376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3035..3054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3245..3266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3385..3405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3390..3719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3570..3705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3632..3651
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3768..4793
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3768..4793
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4923..5044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4960..4979)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5014..5033
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5093..5109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5093..5109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5106..5128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5117..5133
                     /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(5141..5147,5148..5165,5166..5171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5186..5207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5324..5341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5495..6080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5575..5594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6251..7042,7043..7111))
                     /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     6874..6893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7112..7216)
                     /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 gtttaaactt aagcttgcca ccatggctaa gacttccgaa cagagggtga acattgctac
      961 actgctgaca gaaaataaga agaaaatcgt ggataaggct tcccaggatc tgtggcggag
     1021 acacccagac ctgatcgcac caggaggaat tgctttctct cagagggacc gcgctctgtg
     1081 cctgcgagat tacggctggt tcctgcatct gatcaccttt tgtctgctgg ccggagataa
     1141 gggccccatc gagtctattg ggctgatcag tattcgagaa atgtataact cactgggagt
     1201 gcccgtccct gcaatgatgg agagcattag atgcctgaaa gaagccagcc tgtccctgct
     1261 ggacgaagag gacgccaacg agaccgcacc ctactttgat tacattatta aggctatgag
     1321 ctaactcgag tactccggta ttgcggtacc cttgtacgcc tgttttatac tcccttcccg
     1381 taacttagac gcacaaaacc aagttcaata gaagggggta caaaccagta ccaccacgaa
     1441 caagcacttc tgtttccccg gtgatgtcgt atagactgct tgcgtggttg aaagcgacgg
     1501 atccgttatc cgcttatgta cttcgagaag cccagtacca cctcggaatc ttcgatgcgt
     1561 tgcgctcagc actcaacccc agagtgtagc ttaggctgat gagtctggac atccctcacc
     1621 ggtgacggtg gtccaggctg cgttggcggc ctacctatgg ctaacgccat gggacgctag
     1681 ttgtgaacaa ggtgtgaaga gcctattgag ctacataaga atcctccggc ccctgaatgc
     1741 ggctaatccc aacctcggag caggtggtca caaaccagtg attggcctgt cgtaacgcgc
     1801 aagtccgtgg cggaaccgac tactttgggt gtccgtgttt ccttttattt tattgtggct
     1861 gcttatggtg acaatcacag attgttatca taaagcgaat tggataggat ccgtacgatg
     1921 agtgtcaact tagcttccca gttgcgggaa gggacgaaaa aatcccactc catggcggag
     1981 aacgtcggct ttgtcaaatg cttcctcaag ggcgttgtcg agaaaaattc ctaccgtaag
     2041 ctggttggca atctctactt tgtctacagt gccatggaag aggaaatggc aaaatttaag
     2101 gaccatccca tcctcagcca catttacttc cccgaactca accgcaaaca aagcctagag
     2161 caagacctgc aattctatta cggctccaac tggcggcaag aagtgaaaat ttctgccgct
     2221 ggccaagcct atgtggaccg agtccggcaa gtggccgcta cggcccctga attgttggtg
     2281 gcccattcct acacccgtta cctgggggat ctttccggcg gtcaaattct caagaaaatt
     2341 gcccaaaatg ccatgaatct ccacgatggt ggcacagctt tctatgaatt tgccgacatt
     2401 gatgacgaaa aggcttttaa aaatacctac cgtcaagcta tgaatgatct gcccattgac
     2461 caagccaccg ccgaacggat tgtggatgaa gccaatgacg cctttgccat gaacatgaaa
     2521 atgttcaacg aacttgaagg caacctgatc aaggcgatcg gcattatggt gttcaacagc
     2581 ctcacccgtc gccgcagtca aggcagcacc gaagttggcc tcgccacctc cgaaggctag
     2641 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     2701 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2761 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2821 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2881 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     2941 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     3001 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     3061 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3121 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3181 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3241 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3301 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3361 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3421 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3481 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3541 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3601 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     3661 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     3721 agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgatg aaaaagcctg
     3781 aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc
     3841 tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg
     3901 gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc
     3961 ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg gaattcagcg
     4021 agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg
     4081 aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg
     4141 ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca
     4201 ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg
     4261 tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg
     4321 ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc
     4381 tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt
     4441 cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc
     4501 agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt
     4561 atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg
     4621 atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg
     4681 ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt gtagaagtac
     4741 tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa tagcacgtgc
     4801 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4861 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4921 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4981 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5041 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5101 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5161 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5221 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5281 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5341 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5401 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5461 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5521 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5581 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5641 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5701 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5761 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5821 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5881 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5941 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6001 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
     6061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
