LOCUS       sequence-452486-        3764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mTFP1 and extra insert, for MuSIC barcode 
            construction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3764)
  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 3764)
  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-mTFP1
FEATURES             Location/Qualifiers
     source          1..3764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    418..539
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      593..1181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1082..1101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1356..1659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1660..1863
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1814..1834
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1890..1946
                     /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             1956..2009
                     /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      2019..2028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2025..2735
                     /codon_start=1
                     /product="monomeric teal (cyan) fluorescent protein (Ai et 
                     al., 2006)"
                     /label=mTFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEETTMGVIKPDMKIKLKMEGNVNGHAFVIEGEGEGKPYDG
                     TNTINLEVKEGAPLPFSYDILTTAFAYGNRAFTKYPDDIPNYFKQSFPEGYSWERTMTF
                     EDKGIVKVKSDISMEEDSFIYEIHLKGENFPPNGPVMQKKTTGWDASTERMYVRDGVLK
                     GDVKHKLLLEGGGHHRVDFKTIYRAKKAVKLPDYHFVDHRIEILNHDKDYNKVTVYESA
                     VARNSTDGMDELYK"
     polyA_signal    2770..2891
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2898..3353)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3035..3056)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3247..3266
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3379..3481
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3482..3764,1..377)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3548..3567)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc
       61 agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc
      121 ctaaagggtt tattgagaat atgtttttcg tatcagccaa tccctgggtg agtttcacca
      181 gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca
      241 aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg
      301 tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt
      361 ggcagggcgg ggcgtaatca aacatgagaa ttggtcgcgc gagcaattca gacatgataa
      421 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      481 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
      541 gcatttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      601 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      661 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      721 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      781 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      841 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      901 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      961 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     1021 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     1081 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     1141 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1201 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1261 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     1321 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     1381 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     1441 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     1501 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     1561 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     1621 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     1681 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     1741 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     1801 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     1861 gctggtttag tgaagaagac ggctccggcg ccacaaactt ctctctgcta aagcaagcag
     1921 gtgatgttga agaaaacccc gggcctggca gcggcgaggg caggggaagt ctacttacat
     1981 gcggggacgt ggaggaaaat cccggcccag gatccgccgc caccatggtg agcaagggcg
     2041 aggagactac aatgggcgta atcaagcccg acatgaagat caagctgaag atggagggca
     2101 acgtgaatgg ccacgccttc gtgatcgagg gcgagggcga gggcaagccc tacgacggca
     2161 ccaacaccat caacctggag gtgaaggagg gagcccccct gcccttctcc tacgacattc
     2221 tgaccaccgc gttcgcctac ggcaacaggg ccttcaccaa gtaccccgac gacatcccca
     2281 actacttcaa gcagtccttc cccgagggct actcttggga gcgcaccatg accttcgagg
     2341 acaagggcat cgtgaaggtg aagtccgaca tctccatgga ggaggactcc ttcatctacg
     2401 agatacacct caagggcgag aacttccccc ccaacggccc cgtgatgcag aagaaaacca
     2461 ccggctggga cgcctccacc gagaggatgt acgtgcgcga cggcgtgctg aagggcgacg
     2521 tcaagcacaa gctgctgctg gagggcggcg gccaccaccg cgttgacttc aagaccatct
     2581 acagggccaa gaaggcggtg aagctgcccg actatcactt tgtggaccac cgcatcgaga
     2641 tcctgaacca cgacaaggac tacaacaagg tgaccgttta cgagagcgcc gtggcccgca
     2701 actccaccga cggcatggac gagctgtaca agtaaagctt cttattcatg tcttctgaaa
     2761 cataaaatga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2821 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2881 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2941 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3001 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3061 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3121 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3181 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3241 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3301 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcagtact
     3361 taacggaaga tcacttcgtg atcggcacgt aagaggttcc aactttcacc ataatgaaat
     3421 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     3481 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     3541 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     3601 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     3661 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     3721 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttt
//
