LOCUS       sequence-317638-        5055 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Inducible expression of mKate2 under the ZF6x6-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 Apr 14, 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: pJM585 ZF6x6-C mKate2 in TUPV1
FEATURES             Location/Qualifiers
     source          1..5055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      424..852
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(511..530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     721..742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        875..1071
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      922..1057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     984..1003
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1191..1312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1228..1247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1282..1301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1361..1377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1361..1377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1374..1396)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1385..1401
                     /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(1409..1415,1416..1433,1434..1439))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1454..1475
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1592..1609)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1763..2351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1843..1862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2522..3313,3314..3382))
                     /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     3145..3164
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3383..3487)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      3731..3740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3737..4435
                     /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(4480..4497)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4486..4710
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gcgtacggat ggggctctag ctctagagct tgatatcgaa ttcctgcagc cccgctagag
       61 ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg ctagggggca
      121 gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc ccgagccggc
      181 agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc ctctgaacgc
      241 ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa gctttaggct
      301 gaaagagaga tttagaatga cagaatcata gaacggcctg ggcgcgcctg gtggcgaacg
      361 atcaacagac gggagagggc aaaagtcttc gctgacagcg accggactct agggggtatc
      421 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      841 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggc tcaattagtc agcaaccata
      901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     1021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     1081 ggagcttgta tatccatttt cggaggttgg gcttcggaat cgttttccgg gacgccggct
     1141 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1201 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1261 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1321 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1381 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1441 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1501 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1561 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1621 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1681 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1741 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1801 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1861 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1921 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1981 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2041 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2101 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2161 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2221 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2281 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2341 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2401 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2461 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2521 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2581 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2641 ccagtgctgc aatgataccg cgagatccac gctcaccggc tccagattta tcagcaataa
     2701 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2761 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2821 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2881 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2941 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3001 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3061 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3121 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3181 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3241 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3301 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3361 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3421 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3481 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcgaagac
     3541 ggtgccaagg tcttgacaaa cgagtgctag tacaagatct gcgaggtgta ggggacgcga
     3601 gtgtaggggt gagctgtgta gggggactaa gtgtagggga cgcgagtgta ggggtgagct
     3661 gtgtagggga acgctctcta gagggtatat aatgggggcc actagtctac taccagagct
     3721 catcgctagc gccaccatgg tgagcgagct gattaaggag aacatgcaca tgaagctgta
     3781 tatggagggc accgtgaaca accaccactt caagtgtacc tccgagggcg aaggcaagcc
     3841 ctacgagggc acccagacca tgagaatcaa ggcggtcgag ggcggccctc tccccttcgc
     3901 cttcgacatc ctggctacca gcttcatgta cggcagcaaa accttcatca accacaccca
     3961 gggcatcccc gacttcttta agcagtcctt ccccgagggc ttcacatggg agagagtcac
     4021 cacatacgaa gatgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg
     4081 cctcatctac aacgtcaaga tcagaggggt gaacttccca tccaacggcc ctgtgatgca
     4141 gaagaaaaca ctcggctggg aggcctccac cgaaaccctg taccccgctg acggcggcct
     4201 ggaaggcaga gccgacatgg ccctgaagct cgtgggcggg ggccacctga tctgcaactt
     4261 gaaaaccaca tacagatcca agaaacccgc taagaacctc aagatgcccg gcgtctacta
     4321 tgtggacagg agactggaaa gaatcaagga ggccgacaaa gaaacctacg tcgagcagca
     4381 cgaggtggct gtggccagat actgcgacct ccctagcaaa ctggggcaca gatgagcggc
     4441 cgctcgagtc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     4501 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     4561 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     4621 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gtcaatagca
     4681 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc aggtcgaggg tacatgcgcc
     4741 ttactccttg tgagcgcatc gcgataagct ctagagggac agcccccccc caaagccccc
     4801 agggatgtaa ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc
     4861 tccggtccgg cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac
     4921 ggggaaggtg gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca
     4981 gacacctggg gggatacggg gaaaaagctt taggctgaaa gagagattta gaatgacaga
     5041 atcatagaac ggcct
//
