LOCUS       sequence-177144-        5128 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pCAG promoter driving iRFP720.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5128)
  AUTHORS   Weinberg BH, Pham NT, Caraballo LD, Lozanoski T, Engel A, Bhatia S, 
            Wong WW
  TITLE     Large-scale design of robust genetic circuits with multiple inputs 
            and outputs for mammalian cells.
  JOURNAL   Nat Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805.
  PUBMED    28346402
REFERENCE   2  (bases 1 to 5128)
  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 
            Biotechnol. 2017 Mar 27. doi: 10.1038/nbt.3805."
COMMENT     Sequence Label: pCAG-iRFP720
FEATURES             Location/Qualifiers
     source          1..5128
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      626..635
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             632..1582
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     primer_bind     complement(1668..1687)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1733..1788
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1787..1806)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2149..2165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2162..2184)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2173..2189
                     /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(2197..2203,2204..2221,2222..2227))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2242..2263
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2348..2482
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2502..2636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2539..2558)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2593..2612
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2704..2721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2875..3463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2955..2974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3634..4425,4426..4494))
                     /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     4257..4276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4495..4599)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4630..5009
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga acgcgtaccg gtgccgccac catggcggaa ggatccgtcg ccaggcagcc
      661 tgacctcttg acctgcgacg atgagccgat ccatatcccc ggtgccatcc aaccgcatgg
      721 actgctgctc gccctcgccg ccgacatgac gatcgttgcc ggcagcgaca accttcccga
      781 actcaccgga ctggcgatcg gcgccctgat cggccgctct gcggccgatg tcttcgactc
      841 ggagacgcac aaccgtctga cgatcgcctt ggccgagccc ggggcggccg tcggagcacc
      901 gatcactgtc ggcttcacga tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca
      961 tgatcagctc atcttcctcg agctcgagcc tccccagcgg gacgtcgccg agccgcaggc
     1021 gttcttccgc cgcaccaaca gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag
     1081 cgcctgcgcc gccgcggcgc aagaggtgcg gaagattacc ggcttcgatc gggtgatgat
     1141 ctatcgcttc gcctccgact tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt
     1201 cgagtcaaaa ctaggcctgc actatcctgc ctcattcatc ccggcgcagg cccgtcggct
     1261 ctataccatc aacccggtac ggatcattcc cgatatcaat tatcggccgg tgccggtcac
     1321 cccagacctc aatccggtca ccgggcggcc gattgatctt agcttcgcca tcctgcgcag
     1381 cgtctcgccc aaccatctgg agttcatgcg caacataggc atgcacggca cgatgtcgat
     1441 ctcgattttg cgcggcgagc gactgtgggg attgatcgtt tgccatcacc gaacgccgta
     1501 ctacgtcgat ctcgatggcc gccaagcctg cgagctagtc gcccaggttc tggcctggca
     1561 gatcggcgtg atggaagagt gagcggccgc actcctcagg tgcaggctgc ctatcagaag
     1621 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     1681 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     1741 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     1801 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     1861 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     1921 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     1981 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     2041 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     2101 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     2161 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2221 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2281 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     2341 accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2401 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2461 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     2521 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2581 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     2641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4621 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     4681 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4741 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4801 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4861 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4921 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4981 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     5041 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     5101 gtgcagcgat gggggcgggg gggggggg
//
