LOCUS       sequence-452484-        3758 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CyOFP1 and extra insert, for MuSIC barcode 
            construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3758)
  AUTHORS   Lu X, Pritko DJ, Abravanel ME, Huggins JR, Ogunleye O, Biswas T, 
            Ashy KC, Woods SK, Livingston MWT, Blenner MA, Birtwistle MR
  TITLE     Genetically Encoded Fluorescence Barcodes Allow for Single-Cell 
            Analysis via Spectral Flow Cytometry.
  JOURNAL   ACS Synth Biol. 2025 May 16;14(5):1533-1548. doi: 
            10.1021/acssynbio.4c00807. Epub 2025 May 6.
  PUBMED    40326708
REFERENCE   2  (bases 1 to 3758)
  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"; doi: 
            "10.1021/acssynbio.4c00807"; journalName: "ACS Synth Biol"; date: 
            "2025-05-16- 16"; volume: "14"; issue: "5"; pages: "1533-1548"
COMMENT     Sequence Label: pM-CyOFP1
FEATURES             Location/Qualifiers
     source          1..3758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..57
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             113..169
                     /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             179..232
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      242..251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             248..949
                     /codon_start=1
                     /product="orange-red fluorescent protein that is excitable 
                     by cyan light and suitable as an acceptor for blue-emitting
                     luciferase (Chu et al., 2016)"
                     /label=CyOFP1
                     /translation="MVSKGEELIKENMRSKLYLEGSVNGHQFKCTHEGEGKPYEGKQTN
                     RIKVVEGGPLPFAFDILATHFMYGSKVFIKYPADLPDYFKQSFPEGFTWERVMVFEDGG
                     VLTATQDTSLQDGELIYNVKVRGVNFPANGPVMQKKTLGWEPSTETMYPADGGLEGRCD
                     KALKLVGGGHLHVNFKTTYKSKKPVKMPGVHYVDRRLERIKEADNETYVEQYEHAVARY
                     SNLGGGMDELYK"
     polyA_signal    987..1108
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(1115..1570)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1252..1273)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1464..1483
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1596..1698
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1699..2358
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1765..1784)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     polyA_signal    2399..2520
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      2574..3162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3063..3082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3337..3640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(3641..3758,1..86)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
       61 ggtgggaggt ctatataagc agagctggtt tagtgaagaa gacggctccg gcgccacaaa
      121 cttctctctg ctaaagcaag caggtgatgt tgaagaaaac cccgggcctg gcagcggcga
      181 gggcagggga agtctactta catgcgggga cgtggaggaa aatcccggcc caggatccgc
      241 cgccaccatg gtgagcaagg gcgaggagct gatcaaggag aacatgagaa gcaagctgta
      301 cctggaaggc agcgtgaacg gccaccagtt caagtgcacc cacgaagggg agggcaagcc
      361 ctacgagggc aagcagacca acaggatcaa ggtggtggag ggaggccccc tgccgttcgc
      421 attcgacatc ctggccaccc actttatgta cgggagcaag gtgttcatca agtaccccgc
      481 cgacctcccc gattatttta agcagtcctt ccctgagggc ttcacatggg agagagtcat
      541 ggtgttcgag gacgggggcg tgctgaccgc cacccaggac accagcctcc aggacggcga
      601 gctcatctac aacgtcaagg tcagaggggt gaacttccca gccaacggcc ccgtgatgca
      661 gaagaaaaca ctgggctggg aaccaagcac cgaaaccatg tacccagctg acggaggtct
      721 ggaaggaaga tgcgacaagg ccctgaagct cgtgggaggt ggacacttac acgtcaactt
      781 caagaccaca tacaagtcca agaaacccgt gaagatgccc ggcgtccact acgtggaccg
      841 cagactggaa agaatcaagg aggccgacaa cgagacttac gtcgagcagt acgagcacgc
      901 tgtggccaga tactccaacc tgggtggagg catggacgag ctgtacaagt aagagttgcc
      961 attcatgtct tctgaaacat aaaatgaact tgtttattgc agcttataat ggttacaaat
     1021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1081 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     1141 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1201 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     1261 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     1321 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     1381 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     1441 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     1501 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     1561 cagggcgcgt cagtacttaa cggaagatca cttcgtgatc ggcacgtaag aggttccaac
     1621 tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc gagattttca
     1681 ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc
     1741 caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac
     1801 cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag
     1861 ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt
     1921 atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt
     1981 ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg
     2041 cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc
     2101 cctaaagggt ttattgagaa tatgtttttc gtatcagcca atccctgggt gagtttcacc
     2161 agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc
     2221 aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc
     2281 gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag
     2341 tggcagggcg gggcgtaatc aaacatgaga attggtcgcg cgagcaattc agacatgata
     2401 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2461 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2521 ggcatttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3241 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     3301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacggg
//
