LOCUS       sequence-317918-        5062 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Inducible expression of mKate2 under the ZF(2/6)x3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5062)
  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 5062)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pJM597 ZF(2/6)x3 mKate2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      463..891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(550..569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     760..781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        914..1110
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      961..1096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1023..1042
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1230..1351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1267..1286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1321..1340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1400..1416)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1400..1416)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1413..1435)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1424..1440
                     /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(1448..1454,1455..1472,1473..1478))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1493..1514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1631..1648)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1802..2390)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1882..1901)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2561..3352,3353..3421))
                     /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     3184..3203
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3422..3526)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3777..3786
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3783..4481
                     /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(4526..4543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4532..4756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gaaagagaga tttagaatga cagaatcata gaacggcctg cgtacggatg gggctctagc
       61 tctagagctt gatatcgaat tcctgcagcc ccgctagagg gacagccccc ccccaaagcc
      121 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
      181 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
      241 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
      301 gcagacacct ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac
      361 agaatcatag aacggcctgg gcgcgcctgg tggcgaacga tcaacagacg ggagagggca
      421 aaagtcttcg ctgacagcga ccggactcta gggggtatcc ccacgcgccc tgtagcggcg
      481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      901 gaatgtgtgt cagttaggct caattagtca gcaaccatag tcccgcccct aactccgccc
      961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     1021 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     1081 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     1141 ggaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     1201 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     1261 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     1321 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     1381 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1441 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1501 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1561 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1621 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1681 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1741 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1801 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1861 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1921 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1981 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2041 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2101 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2161 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2221 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2281 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2341 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2401 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2461 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2521 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2581 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2641 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2701 gagatccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2761 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2821 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2881 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2941 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3001 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3061 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3121 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3181 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3241 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3301 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3361 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3421 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3481 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3541 aagtgccacc tgacgtcgac ggatcgggag atcgaagacg gtgccaaggt cttgacaaac
     3601 gagtgctagt acaagatctg aattacgatc tgaggacgtg taattggtgt aggggtgatc
     3661 tgaggacgtg taattggtgt aggggtgatc tgaggacgtg taattggtgt aggggtgatc
     3721 cctctagagg gtatataatg ggggccacta gtctactacc agagctcatc gctagcgcca
     3781 ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtatatg gagggcaccg
     3841 tgaacaacca ccacttcaag tgtacctccg agggcgaagg caagccctac gagggcaccc
     3901 agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
     3961 ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact
     4021 tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca tacgaagatg
     4081 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     4141 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     4201 gctgggaggc ctccaccgaa accctgtacc ccgctgacgg cggcctggaa ggcagagccg
     4261 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaaa accacataca
     4321 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacaggagac
     4381 tggaaagaat caaggaggcc gacaaagaaa cctacgtcga gcagcacgag gtggctgtgg
     4441 ccagatactg cgacctccct agcaaactgg ggcacagatg agcggccgct cgagtctaga
     4501 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4561 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4621 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4681 ggggtggggc aggacagcaa gggggaggat tgggaagtca atagcaggca tgctggggat
     4741 gcggtgggct ctatggcttc tgaggcaggt cgagggtaca tgcgccttac tccttgtgag
     4801 cgcatcgcga taagctctag agggacagcc cccccccaaa gcccccaggg atgtaattac
     4861 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     4921 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     4981 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     5041 tacggggaaa aagctttagg ct
//
