LOCUS       sequence-317642-        5086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of mKate2 under the ZF1x6-C promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5086)
  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 5086)
  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: pJM410 ZF1x6-C mKate2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      698..1126
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(785..804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     995..1016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1149..1345
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1196..1331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1258..1277
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1465..1586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1502..1521)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1556..1575
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1635..1651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1635..1651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1648..1670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1659..1675
                     /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(1683..1689,1690..1707,1708..1713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1728..1749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1866..1883)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2037..2625)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2117..2136)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2796..3587,3588..3656))
                     /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     3419..3438
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3657..3761)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      4036..4045
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4042..4740
                     /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(4785..4802)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4791..5015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc
       61 gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt
      121 gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc
      181 gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt aggctgaaag
      241 agagatttag aatgacagaa tcatagaacg gcctgcgtac ggatggggct ctagctctag
      301 agcttgatat cgaattcctg cagccccgct agagggacag ccccccccca aagcccccag
      361 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
      421 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
      481 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
      541 cacctggggg gatacgggga aaaagcttta ggctgaaaga gagatttaga atgacagaat
      601 catagaacgg cctgggcgcg cctggtggcg aacgatcaac agacgggaga gggcaaaagt
      661 cttcgctgac agcgaccgga ctctaggggg tatccccacg cgccctgtag cggcgcatta
      721 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
      781 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
      841 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
      901 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
      961 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1021 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1081 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     1141 tgtgtcagtt aggctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1201 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1261 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     1321 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggagg
     1381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1441 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     1501 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1561 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     1621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1801 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1861 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1921 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1981 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2041 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2101 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2161 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2221 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2281 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2341 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2401 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2461 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2521 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2581 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2641 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2701 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2761 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2821 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     2881 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagat
     2941 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3001 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3061 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3121 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3181 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3241 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3301 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3361 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3421 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3481 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3541 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3601 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3781 ccacctgacg tcgacggatc gggagatcga agacggtgcc aaggtcttga caaacgagtg
     3841 ctagtacaag atcttgacgt cgacggatcg ggagatctga attacgatcg gagtgaggat
     3901 aattagagtg aggacgatcg gagtgaggaa aattagagtg aggacgatcg gagtgaggat
     3961 aattagagtg aggacgatcc ctctagaggg tatataatgg gggccactag tctactacca
     4021 gagctcatcg ctagcgccac catggtgagc gagctgatta aggagaacat gcacatgaag
     4081 ctgtatatgg agggcaccgt gaacaaccac cacttcaagt gtacctccga gggcgaaggc
     4141 aagccctacg agggcaccca gaccatgaga atcaaggcgg tcgagggcgg ccctctcccc
     4201 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac
     4261 acccagggca tccccgactt ctttaagcag tccttccccg agggcttcac atgggagaga
     4321 gtcaccacat acgaagatgg gggcgtgctg accgctaccc aggacaccag cctccaggac
     4381 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     4441 atgcagaaga aaacactcgg ctgggaggcc tccaccgaaa ccctgtaccc cgctgacggc
     4501 ggcctggaag gcagagccga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     4561 aacttgaaaa ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     4621 tactatgtgg acaggagact ggaaagaatc aaggaggccg acaaagaaac ctacgtcgag
     4681 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacagatga
     4741 gcggccgctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     4801 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     4861 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     4921 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagtcaa
     4981 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcaggtc gagggtacat
     5041 gcgccttact ccttgtgagc gcatcgcgat aagctctaga gggaca
//
