LOCUS       sequence-151193-        7715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Tandem Dimer smURFP and Heme Oxygenase-1 in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7715)
  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 7715)
  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-TDsmURFP-IRES-HO-1
FEATURES             Location/Qualifiers
     source          1..7715
                     /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..1792
                     /codon_start=1
                     /product="monomeric tandem dimer of a small ultra-red 
                     fluorescent protein derived from a cyanobacterial 
                     phycobiliprotein (Rodriguez et al., 2016)"
                     /label=TDsmURFP
                     /translation="MAKTSEQRVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAF
                     SQRDRALCLRDYGWFLHLITFCLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRC
                     LKEASLSLLDEEDANETAPYFDYIIKAMSGHGTGSTGSGSSGTASSEDNNMAMAKTSEQ
                     RVNIATLLTENKKKIVDKASQDLWRRHPDLIAPGGIAFSQRDRALCLRDYGWFLHLITF
                     CLLAGDKGPIESIGLISIREMYNSLGVPVPAMMESIRCLKEASLSLLDEEDANETAPYF
                     DYIIKAMS"
     primer_bind     complement(3140..3157)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3146..3370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3416..3844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3713..3734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3853..3873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3858..4187
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4038..4173
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4100..4119
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4236..5261
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4236..5261
                     /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    5391..5512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5428..5447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5482..5501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5561..5577)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5561..5577)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5574..5596)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5585..5601
                     /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(5609..5615,5616..5633,5634..5639))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5654..5675
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5792..5809)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5963..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6043..6062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6719..7510,7511..7579))
                     /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     7342..7361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7580..7684)
                     /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 cgggcatggc accggcagca ccggcagcgg cagctccggc accgcctcct ccgaggacaa
     1381 caacatggcc atggctaaga cttccgaaca gagggtgaac attgctacac tgctgacaga
     1441 aaataagaag aaaatcgtgg ataaggcttc ccaggatctg tggcggagac acccagacct
     1501 gatcgcacca ggaggaattg ctttctctca gagggaccgc gctctgtgcc tgcgagatta
     1561 cggctggttc ctgcatctga tcaccttttg tctgctggcc ggagataagg gccccatcga
     1621 gtctattggg ctgatcagta ttcgagaaat gtataactca ctgggagtgc ccgtccctgc
     1681 aatgatggag agcattagat gcctgaaaga agccagcctg tccctgctgg acgaagagga
     1741 cgccaacgag accgcaccct actttgatta cattattaag gctatgagct aactcgagta
     1801 ctccggtatt gcggtaccct tgtacgcctg ttttatactc ccttcccgta acttagacgc
     1861 acaaaaccaa gttcaataga agggggtaca aaccagtacc accacgaaca agcacttctg
     1921 tttccccggt gatgtcgtat agactgcttg cgtggttgaa agcgacggat ccgttatccg
     1981 cttatgtact tcgagaagcc cagtaccacc tcggaatctt cgatgcgttg cgctcagcac
     2041 tcaaccccag agtgtagctt aggctgatga gtctggacat ccctcaccgg tgacggtggt
     2101 ccaggctgcg ttggcggcct acctatggct aacgccatgg gacgctagtt gtgaacaagg
     2161 tgtgaagagc ctattgagct acataagaat cctccggccc ctgaatgcgg ctaatcccaa
     2221 cctcggagca ggtggtcaca aaccagtgat tggcctgtcg taacgcgcaa gtccgtggcg
     2281 gaaccgacta ctttgggtgt ccgtgtttcc ttttatttta ttgtggctgc ttatggtgac
     2341 aatcacagat tgttatcata aagcgaattg gataggatcc gtacgatgag tgtcaactta
     2401 gcttcccagt tgcgggaagg gacgaaaaaa tcccactcca tggcggagaa cgtcggcttt
     2461 gtcaaatgct tcctcaaggg cgttgtcgag aaaaattcct accgtaagct ggttggcaat
     2521 ctctactttg tctacagtgc catggaagag gaaatggcaa aatttaagga ccatcccatc
     2581 ctcagccaca tttacttccc cgaactcaac cgcaaacaaa gcctagagca agacctgcaa
     2641 ttctattacg gctccaactg gcggcaagaa gtgaaaattt ctgccgctgg ccaagcctat
     2701 gtggaccgag tccggcaagt ggccgctacg gcccctgaat tgttggtggc ccattcctac
     2761 acccgttacc tgggggatct ttccggcggt caaattctca agaaaattgc ccaaaatgcc
     2821 atgaatctcc acgatggtgg cacagctttc tatgaatttg ccgacattga tgacgaaaag
     2881 gcttttaaaa atacctaccg tcaagctatg aatgatctgc ccattgacca agccaccgcc
     2941 gaacggattg tggatgaagc caatgacgcc tttgccatga acatgaaaat gttcaacgaa
     3001 cttgaaggca acctgatcaa ggcgatcggc attatggtgt tcaacagcct cacccgtcgc
     3061 cgcagtcaag gcagcaccga agttggcctc gccacctccg aaggctagtc tagagggccc
     3121 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3181 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3241 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3301 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3361 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3541 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3781 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3841 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3961 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4021 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4081 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4141 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4201 cttgtatatc cattttcgga tctgatcagc acgtgatgaa aaagcctgaa ctcaccgcga
     4261 cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct
     4321 cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc
     4381 gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat
     4441 cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct
     4501 attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc
     4561 ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     4621 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     4681 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     4741 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     4801 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     4861 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     4921 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     4981 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     5041 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     5101 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     5161 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     5221 gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtgcta cgagatttcg
     5281 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5341 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5401 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5461 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5521 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5581 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5641 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5701 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5761 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5821 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5881 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5941 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6001 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6061 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6121 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6181 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6241 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6301 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6361 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6421 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6481 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6541 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6601 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6661 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6721 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6781 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6841 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6901 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6961 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7021 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7081 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7141 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7201 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7261 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7321 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7381 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7441 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7501 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7561 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7621 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7681 cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
