LOCUS       2xMyc-tdMCP-SNAP        6455 bp DNA     circular SYN 02-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6455)
  AUTHORS   Zack
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2024 from SnapGene 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(404..1195,1196..1264))
                     /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"
     promoter        complement(1265..1369)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1635..2014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2015..2218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    2220..2238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2241..2259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2396..2425
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2435..2464
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2465..2485
                     /codon_start=1
                     /label=linker
                     /translation="GSGSGSG"
     CDS             2486..2833
                     /codon_start=1
                     /label=MCP (codon optimized #1)
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGIAEWISSNSRSQAYKVT
                     CSVRQSSAQNRKYTIKVEVPKGAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGNP
                     IPSAIAANSGIY"
     CDS             2837..3187
                     /codon_start=1
                     /label=MCP (codon optimized #2)
                     /translation="MASNFTQFVLVDNGGTGDVTVAPSNFANGIAEWISSNSRSQAYKV
                     TCSVRQSSAQNRKYTIKVEVPKGAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGN
                     PIPSAIAANSGIY"
     CDS             3188..3205
                     /codon_start=1
                     /label=Linker (codon opt)
                     /translation="GSGGSG"
     CDS             3206..3220
                     /codon_start=1
                     /label=extended linker
                     /translation="SGSGG"
     CDS             3221..3766
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     polyA_signal    3813..4037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4321..4368
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4376..5398
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5528..5649
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5698..5714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5722..5738
                     /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(5746..5752,5753..5770,5771..5776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5791..5812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(join(6100..6455,1..233))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
       61 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1381 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     1441 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     1501 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     1561 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     1621 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1681 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1741 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1801 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1861 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1921 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1981 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     2041 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2101 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     2161 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
     2221 ccctatcagt gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt
     2281 gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg
     2341 ggaccgatcc agcctccgga ctctagcgtt taaacttaag cttgccgcca ccatggagca
     2401 aaaactcatc agcgaggaag atcttgggtc aggagagcag aaattgattt cagaggaaga
     2461 cttgggttca ggaagcggtt ctggagcctc caattttact cagttcgtct tggtggataa
     2521 cggcggtacg ggggacgtta cggtcgctcc ctccaatttt gccaacggaa tagccgagtg
     2581 gatttccagt aattccaggt cacaagcgta taaagtaacg tgttccgtca gacaaagctc
     2641 cgcccaaaat cgaaagtata ccatcaaagt agaagttcca aagggtgcgt ggagatctta
     2701 tctgaacatg gaactcacta ttcccatctt cgccaccaac tccgactgtg aacttatcgt
     2761 taaggcaatg caaggtctgc tgaaagatgg aaaccctatc cctagcgcca tcgccgcaaa
     2821 ctccggcatc tatgccatgg cttcaaactt cacccagttt gtgcttgtag acaatggtgg
     2881 cacgggcgac gtgacagtgg caccttccaa ctttgctaac ggcattgcag aatggattag
     2941 ttctaattcc cgctcccagg cttataaagt tacgtgtagt gttcgccagt caagcgcaca
     3001 aaaccgaaag tacaccataa aggttgaagt ccccaaggga gcctggcgat cataccttaa
     3061 catggagctt actattccga tcttcgctac caattcagat tgtgaattga ttgtcaaagc
     3121 tatgcaaggc ctgctcaagg atggtaaccc gatacctagc gctatcgccg ccaattcagg
     3181 catttatggc agtggaggtt ccggatccgg atctggtggc atggacaaag actgcgaaat
     3241 gaagcgcacc accctggata gccctctggg caagctggaa ctgtctgggt gcgaacaggg
     3301 cctgcaccgt atcatcttcc tgggcaaagg aacatctgcc gccgacgccg tggaagtgcc
     3361 tgccccagcc gccgtgctgg gcggaccaga gccactgatg caggccaccg cctggctcaa
     3421 cgcctacttt caccagcctg aggccatcga ggagttccct gtgccagccc tgcaccaccc
     3481 agtgttccag caggagagct ttacccgcca ggtgctgtgg aaactgctga aagtggtgaa
     3541 gttcggagag gtcatcagct acagccacct ggccgccctg gccggcaatc ccgccgccac
     3601 cgccgccgtg aaaaccgccc tgagcggaaa tcccgtgccc attctgatcc cctgccaccg
     3661 ggtggtgcag ggcgacctgg acgtgggggg ctacgagggc gggctcgccg tgaaagagtg
     3721 gctgctggcc cacgagggcc acagactggg caagcctggg ctgggttgac tcgagtctag
     3781 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     3841 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     3901 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     3961 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4021 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     4081 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4141 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4201 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4261 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta
     4321 gaagttccta ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa
     4381 gcctgaactc accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc
     4441 cgacctgatg cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg
     4501 gcgtggatat gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt
     4561 ttatcggcac tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt
     4621 cagcgagagc ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct
     4681 gcctgaaacc gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc
     4741 tgcggccgat cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca
     4801 atacactaca tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca
     4861 aactgtgatg gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct
     4921 ttgggccgag gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa
     4981 tgtcctgacg gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg
     5041 ggattcccaa tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga
     5101 gcagcagacg cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg
     5161 ggcgtatatg ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt
     5221 cgatgatgca gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac
     5281 tgtcgggcgt acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga
     5341 agtactcgcc gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca
     5401 cgtactacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     5461 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     5521 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     5581 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     5641 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     5701 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5761 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5821 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6421 ggtaagacac gacttatcgc cactggcagc agcca
//
