LOCUS       sequence-325936-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Split GFP reporter  plasmid expressing GFP (b11beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  AUTHORS   Huang X, Lv J, Li Y, Mao S, Li Z, Jing Z, Sun Y, Zhang X, Shen S, 
            Wang X, Di M, Ge J, Huang X, Zuo E, Chi T
  TITLE     Programmable C-to-U RNA editing using the human APOBEC3A deaminase.
  JOURNAL   EMBO J. 2020 Nov 16;39(22):e104741. doi: 10.15252/embj.2020104741. 
            Epub 2020 Oct 15.
  PUBMED    33058229
REFERENCE   2  (bases 1 to 4740)
  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: "EMBO J."; 
            date: "2020-11-16"; volume: "39(22):e104741. doi"; pages: " 
            10.15252/embj.2020104741. Epub 2020 Oct 15"
COMMENT     Sequence Label: TC1154
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             724..771
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             937..993
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             994..1926
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     misc_feature    1951..2539
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2004..2024)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2422..2433)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2542..2558)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2543..2559)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2560..2577)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2566..2773
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2837..2854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3008..3596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3088..3107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3767..4558,4559..4627))
                     /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     4390..4409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4628..4732)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctatcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcgaatt cgccgccacc
      601 atgaaggtta gcgcactgaa ggaaaaagtt tcagcactga aagagaaggt gagtgccctc
      661 aaagagaaag tctccgctct caaggaaaaa gtctcagccc ttaaagaggg aggatctggc
      721 ggcagggacc acatggtcct gcacgagtac gtcaatgcag caggaatcac cggatcctcg
      781 aatattacga ccctcgaggg cggcagcggc ggcgaagtaa gtgctcttga aaaagaggtg
      841 tctgcccttg aaaaggaagt ctcagcactg gaaaaggagg tatccgcact tgagaaggag
      901 gtaagtgccc tggagaaggc ggccgcgggc agtggggcta ctaacttcag cctgctgaag
      961 caggctggag acgtggagga gaaccctgga cctatggcgc gtaaggtcga tctcacctcc
     1021 tgcgatcgcg agccgatcca catccccggc agcattcagc cgtgcggctg cctgctagcc
     1081 tgcgacgcgc aggcggtgcg gatcacgcgc attacggaaa atgccggcgc gttctttgga
     1141 cgcgaaactc cgcgggtcgg tgagctactc gccgattact tcggcgagac cgaagcccat
     1201 gcgctgcgca acgcactggc gcagtcctcc gatccaaagc gaccggcgct gatcttcggt
     1261 tggcgcgacg gcctgaccgg ccgcaccttc gacatctcac tgcatcgcca tgacggtaca
     1321 tcgatcatcg agttcgagcc tgcggcggcc gaacaggccg acaatccgct gcggctgacg
     1381 cggcagatca tcgcgcgcac caaagaactg aagtcgctcg aagagatggc cgcacgggtg
     1441 ccgcgctatc tgcaggcgat gctcggctat caccgcgtga tgttgtaccg cttcgcggac
     1501 gacggctccg ggatggtgat cggcgaggcg aagcgcagcg acctggagag ctttctcggt
     1561 cagcactttc cggcgtcgct ggtcccgcag caggcgcggc tactgtactt gaagaacgcg
     1621 atccgcgtgg tctcggattc gcgcggcatc agcagccgga tcgtgcccga gcacgacgcc
     1681 tccggcgccg cgctcgatct gtcgttcgcg cacctgcgca gcatctcgcc ctgccatctc
     1741 gaatttctgc ggaacatggg cgtcagcgcc tcgatgtcgc tgtcgatcat cattgacggc
     1801 acgctatggg gattgatcat ctgtcatcat tacgagccgc gtgccgtgcc gatggcgcag
     1861 cgcgtcgcgg ccgaaatgtt cgccgacttc ttatcgctgc acttcaccgc cgcccaccac
     1921 caacgcagca agctgtgata ggctagctag aatcaacctc tggattacaa aatttgtgaa
     1981 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2041 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2101 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2161 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2221 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2281 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2341 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2401 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2461 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2521 ctttgggccg cctccccgca tcgataccgt cgacctcgac ctcgactgtg ccttctagtt
     2581 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     2641 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     2701 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     2761 ggcatgctgg ggagcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2821 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2881 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2941 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3001 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3061 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3121 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3181 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3241 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3301 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3361 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3421 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3481 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3541 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3601 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3661 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3721 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3781 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3841 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3901 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3961 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4021 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4081 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4141 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4201 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4261 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4321 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4381 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4441 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4501 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4561 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4621 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4681 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
//
