LOCUS       sequence-317648-        5055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of mKate2 under the ZF2x6-C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5055)
  AUTHORS   Muldoon JJ, Kandula V, Hong M, Donahue PS, Boucher JD, Bagheri N, 
            Leonard JN
  TITLE     Model-guided design of mammalian genetic programs.
  JOURNAL   Sci Adv. 2021 Feb 19;7(8). pii: 7/8/eabe9375. doi: 
            10.1126/sciadv.abe9375. Print 2021 Feb.
  PUBMED    33608279
REFERENCE   2  (bases 1 to 5055)
  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: "Sci Adv."; 
            date: "2021-02-19"; volume: "7(8). pii: 7/8/eabe9375. doi"; pages: "
            10.1126/sciadv.abe9375. Print 2021 Feb"
COMMENT     Sequence Label: pJM584 ZF2x6-C mKate2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      307..316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             313..1011
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(1056..1073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1062..1286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2055..2483
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2142..2161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2352..2373
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2506..2702
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2553..2688
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2615..2634
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2822..2943
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2859..2878)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2913..2932
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2992..3008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2992..3008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3005..3027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3016..3032
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3040..3046,3047..3064,3065..3070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3085..3106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3223..3240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3394..3982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3474..3493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4153..4944,4945..5013))
                     /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     4776..4795
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5014..5055,1..63))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
       61 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gaagacggtg
      121 ccaaggtctt gacaaacgag tgctagtaca agatctgcga ggaggacgtg acgcgagagg
      181 acgtgtgagc tgaggacgtg gactaagagg acgtgacgcg agaggacgtg tgagctgagg
      241 acgtgaacgc tctctagagg gtatataatg ggggccacta gtctactacc agagctcatc
      301 gctagcgcca ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtatatg
      361 gagggcaccg tgaacaacca ccacttcaag tgtacctccg agggcgaagg caagccctac
      421 gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc
      481 gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc
      541 atccccgact tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca
      601 tacgaagatg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      661 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
      721 aaaacactcg gctgggaggc ctccaccgaa accctgtacc ccgctgacgg cggcctggaa
      781 ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaaa
      841 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
      901 gacaggagac tggaaagaat caaggaggcc gacaaagaaa cctacgtcga gcagcacgag
      961 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagatg agcggccgct
     1021 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     1081 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     1141 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     1201 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca
     1261 tgctggggat gcggtgggct ctatggcttc tgaggcaggt cgagggtaca tgcgccttac
     1321 tccttgtgag cgcatcgcga taagctctag agggacagcc cccccccaaa gcccccaggg
     1381 atgtaattac gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg
     1441 gtccggcgct ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg
     1501 aaggtggcac gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca
     1561 cctgggggga tacggggaaa aagctttagg ctgaaagaga gatttagaat gacagaatca
     1621 tagaacggcc tgcgtacgga tggggctcta gctctagagc ttgatatcga attcctgcag
     1681 ccccgctaga gggacagccc ccccccaaag cccccaggga tgtaattacg tccctccccc
     1741 gctagggggc agcagcgagc cgcccggggc tccgctccgg tccggcgctc cccccgcatc
     1801 cccgagccgg cagcgtgcgg ggacagcccg ggcacgggga aggtggcacg ggatcgcttt
     1861 cctctgaacg cttctcgctg ctctttgagc ctgcagacac ctggggggat acggggaaaa
     1921 agctttaggc tgaaagagag atttagaatg acagaatcat agaacggcct gggcgcgcct
     1981 ggtggcgaac gatcaacaga cgggagaggg caaaagtctt cgctgacagc gaccggactc
     2041 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2101 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2161 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     2221 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2281 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2341 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2401 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2461 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg ctcaattagt
     2521 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2581 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2641 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2701 aaaagctccc gggagcttgt atatccattt tcggaggttg ggcttcggaa tcgttttccg
     2761 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     2821 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     2881 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     2941 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3001 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3061 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3121 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3181 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3361 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4141 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4201 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4261 accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg ctccagattt
     4321 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4381 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4441 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4501 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4561 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4621 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4681 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4741 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4801 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4861 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4921 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4981 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5041 catttatcag ggtta
//
