LOCUS       sequence-249770-        2710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a nuclear localized iRFP670 coexpression cassette as a Type 
            4a part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2710)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 2710)
  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.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK4a_011
FEATURES             Location/Qualifiers
     source          1..2710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      56..644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     545..564
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             768..821
                     /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"
     CDS             828..848
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             864..1796
                     /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="ARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAGA
                     FFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLHR
                     HDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVML
                     YRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSRI
                     VPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEP
                     RAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     terminator      complement(1820..1854)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1925..2584)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2499..2518
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
       61 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      121 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
      241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
      301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
      361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
      421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
      481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
      541 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
      601 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
      661 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
      721 cccctgattc tgtggataac cgtagtcggt ctcaatccgg aagcggagag ggcagaggaa
      781 gtctgctaac atgcggtgac gtcgaggaga atcctggacc tggatccccc aagaagaaga
      841 ggaaggtgga ggacgccgga tccgcgcgta aggtcgatct cacctcctgc gatcgcgagc
      901 cgatccacat ccccggcagc attcagccgt gcggctgcct gctagcctgc gacgcgcagg
      961 cggtgcggat cacgcgcatt acggaaaatg ccggcgcgtt ctttggacgc gaaactccgc
     1021 gggtcggtga gctactcgcc gattactttg gcgaaaccga agcccatgcg ctgcgcaacg
     1081 cactggcgca gtcctccgat ccaaagcgac cggcgctgat cttcggttgg cgcgacggcc
     1141 tgaccggccg caccttcgac atctcactgc atcgccatga cggtacatcg atcatcgagt
     1201 tcgagcctgc ggcggccgaa caggccgaca atccgctgcg gctgacgcgg cagatcatcg
     1261 cgcgcaccaa agaactgaag tcgctcgaag agatggccgc acgggtgccg cgctatctgc
     1321 aggcgatgct cggctatcac cgcgtgatgt tgtaccgctt cgcggacgac ggctccggga
     1381 tggtgatcgg cgaggcgaag cgcagcgacc tggagagctt tctcggtcag cactttccgg
     1441 cgtcgctggt cccgcagcag gcgcggctac tgtacttgaa gaacgcgatc cgcgtggtat
     1501 cggattcgcg cggcatcagc agccggatcg tgcccgagca cgacgcctcc ggcgccgcgc
     1561 tcgatctgtc gttcgcgcac ctgcgcagca tctcgccctg ccatctcgaa tttctgcgga
     1621 acatgggcgt cagcgcctcg atgtcgctgt cgatcatcat tgacggcacg ctatggggat
     1681 tgatcatctg tcatcattac gagccgcgtg ccgtgccgat ggcgcagcgc gtcgcggccg
     1741 aaatgttcgc cgacttctta tcgctgcact tcaccgccgc ccaccaccaa cgctaactcg
     1801 agtggctgag accagaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     1861 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat
     1921 caaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc
     1981 cgacatggaa gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct
     2041 tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat
     2101 tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca
     2161 tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt
     2221 gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa
     2281 acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca
     2341 gctcaccgtc tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa
     2401 tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg
     2461 taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa
     2521 aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct
     2581 ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     2641 atcttatttc attatggtga aagttggaac ctcttacgtg cccgatcaat catgaccaaa
     2701 atcccttaac
//
