LOCUS       sequence-396774-        4785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a secreted version of iRFP720 constitutively in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4785)
  AUTHORS   Gayet RV, Ilia K, Razavi S, Tippens ND, Lalwani MA, Zhang K, Chen 
            JX, Chen JC, Vargas-Asencio J, Collins JJ
  TITLE     Autocatalytic base editing for RNA-responsive translational control.
  JOURNAL   Nat Commun. 2023 Mar 11;14(1):1339. doi: 10.1038/s41467-023-36851-z.
  PUBMED    36906659
REFERENCE   2  (bases 1 to 4785)
  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 
            Commun."; date: "2023-03-11"; volume: "14(1):1339. doi"; pages: " 
            10.1038/s41467-023-36851-z"
COMMENT     Sequence Label: pDV05.2_pEF1a-iRFP720_secreted
FEATURES             Location/Qualifiers
     source          1..4785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(251..839)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(331..350)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1010..1801,1802..1870))
                     /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     1633..1652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1871..1975)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2422..2633
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2634..3572
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3529..3549
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3615..3624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3672..3695
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3711..4658
                     /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="AEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGSD
                     NLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFIG
                     SWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKITG
                     FDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDIN
                     YRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGLI
                     VCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     polyA_signal    4711..4759
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgctggtcta atacccaatc tctcgtctta tccagatgtt ttatacgcct taattaatag
       61 ggataacagg gtaatatctt gccctgcacg aataccagcg accccttgcc caaatacttg
      121 ccgtgggcct cggcctgaga gccaaaacac ttgatgcgga agaagtcggt gcgctcctgc
      181 ttgtcgccgg catcgttgcg ccacatctag gatctgccag gaaccgtaaa aaggccgcgt
      241 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      301 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      361 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      421 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
      481 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
      541 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
      601 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
      661 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
      721 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
      781 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
      841 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
      901 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      961 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1021 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1081 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1141 tgataccgcg agaaccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1201 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1261 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1321 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1381 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1441 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1501 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1561 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1621 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1681 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1741 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1801 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1861 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcaaggt tattgtctca
     1921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacga
     1981 attggccagc gctgcattac cctgttatcc ctaaccggtg cgtttttatg cttgtagtat
     2041 tgtataatgt ttttaagatc cggaggccac ctttggtctt gacattcact aaggagtggg
     2101 cctggaataa aaccagaaat ccccattctc agtctgcccc aagccctcca attccgaggg
     2161 cgctgtatgt ataagctcgg gcaggcaaaa gtcgactgtg agaacacgcc agcagagggc
     2221 gctgtggccc catcagtccc tgcccactga actctccaga aggaaaagcg gcaaggatgc
     2281 aaacaagaaa atcagaccag actgggagtc tagaccctgc gtcccacgcg gtcccaccat
     2341 ggcttcctct ttggaatttt cagggggacc caggaagact gacaccgctg attactttaa
     2401 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgt
     2461 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     2521 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     2581 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     2641 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     2701 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     2761 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     2821 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     2881 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     2941 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     3001 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     3061 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     3121 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     3181 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     3241 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     3301 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     3361 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     3421 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     3481 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     3541 cagtggttca aagttttttt cttccatttc aggtggatcc caagggcaga taagtttgta
     3601 caaaaaagag aggtgccacc atgggcgtga aggtcctctt cgcactcatt tgcatagccg
     3661 tagcagaagc cgactataaa gacgacgatg acaagggcgg gtccggcgga gcggaaggat
     3721 ccgtcgccag gcagcctgac ctcttgacct gcgacgatga gccgatccat atccccggtg
     3781 ccatccaacc gcatggactg ctgctcgccc tcgccgccga catgacgatc gttgccggca
     3841 gcgacaacct tcccgaactc accggactgg cgatcggcgc cctgatcggc cgctctgcgg
     3901 ccgatgtctt cgactcggag acgcacaacc gtctgacgat cgccttggcc gagcccgggg
     3961 cggccgtcgg agcaccgatc actgtcggct tcacgatgcg aaaggacgca ggcttcatcg
     4021 gctcctggca tcgccatgat cagctcatct tcctcgagct cgagcctccc cagcgggacg
     4081 tcgccgagcc gcaggcgttc ttccgccgca ccaacagcgc catccgccgc ctgcaggccg
     4141 ccgaaacctt ggaaagcgcc tgcgccgccg cggcgcaaga ggtgcggaag attaccggct
     4201 tcgatcgggt gatgatctat cgcttcgcct ccgacttcag cgggtccgtg atcgcagagg
     4261 atcggtgcgc cgaggtcgag tcaaaactag gcctgcacta tcctgcctca ttcatcccgg
     4321 cgcaggcccg tcggctctat accatcaacc cggtacggat cattcccgat atcaattatc
     4381 ggccggtgcc ggtcacccca gacctcaatc cggtcaccgg gcggccgatt gatcttagct
     4441 tcgccatcct gcgcagcgtc tcgcccaacc atctggagtt catgcgcaac ataggcatgc
     4501 acggcacgat gtcgatctcg attttgcgcg gcgagcgact gtggggattg atcgtttgcc
     4561 atcaccgaac gccgtactac gtcgatctcg atggccgcca agcctgcgag ctagtcgccc
     4621 aggttctggc ctggcagatc ggcgtgatgg aagagtaagc ttccactgga ttgtacaatt
     4681 accaactgat ctagaaccgg tcatggccgc aataaaatat ctttattttc attacatctg
     4741 tgtgttggtt ttttgtgtgt tcgaactaga tgctgtcgac cgatg
//
