LOCUS       62056_17180        6337 bp DNA     circular SYN 21-APR-2021
DEFINITION  Cloning vector pMMZ272, complete sequence.
ACCESSION   MW731450
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6337)
  AUTHORS   Mansouri M, Hussherr M-D., Strittmatter T, Buchmann P, Xue S, 
            Camenisch G, Fussenegger M.
  TITLE     Smart-Watch-Programmed Green-Light-Operated Percutaneous Control of 
            Therapeutic Transgenes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6337)
  AUTHORS   Mansouri M, Hussherr M-D., Strittmatter T, Buchmann P, Xue S, 
            Camenisch G, Fussenegger M.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-MAR-2021) DBSSE, ETH, Mattenstrasse 26, Basel, Choose 
            A State 4058, Switzerland
REFERENCE   3  (bases 1 to 6337)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-MAR-2021) DBSSE, ETH, Mattenstrasse 26, Basel, Choose A State 
            4058, Switzerland"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(49..65)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(73..89)
                     /label=lac operator
                     /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(97..127)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(142..163)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(451..1036)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1210..2067)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2068..2172)
                     /label=AmpR promoter
     promoter        2494..2512
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3204..3821
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     CDS             3840..4073
                     /codon_start=1
                     /label=VP16 AD
                     /note="transcriptional activation domain of herpes simplex
                     virus protein VP16 (Triezenberg et al., 1988; Cousens et 
                     al., 1989)"
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     polyA_signal    4140..4364
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4410..4838
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4852..5181
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5248..6039
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6216..6337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
       61 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
      121 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
      181 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
      241 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      301 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      361 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      421 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      481 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      541 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      601 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      661 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      721 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      781 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      841 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      901 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      961 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     1021 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1081 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1141 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1201 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1261 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1321 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1381 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1441 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1501 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1561 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1621 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1681 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1741 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1801 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1861 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1921 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1981 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2041 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2101 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2161 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     2221 ccacgcgtgg taccctcgag gcgatctgca tctcaattag tcagcaacca tagtcccgcc
     2281 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatcg
     2341 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     2401 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaagagctc tctggctaac
     2461 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     2521 agctgttcga agcggaattc accatgacta gtcccgagga cctcggcacc ggactcctcg
     2581 aagcactctt gagaggagat ttggccggcg ccgaggctct ctttcgacgg gggcttaggt
     2641 tttggggacc cgagggtatt ctggagcacc tgctcctgcc tgtgcttcgg gaggtgggag
     2701 aagcctggca tcgcggcgag atcggtgtgg ccgaggagca tctggcatcc acatttctgc
     2761 gcgcgagact gcaggagctg ctcgacctcg ccgggttccc acctggcccc cccgtgctgg
     2821 taaccacgcc accaggggag aggcacgaga tcggcgcaat gttggctgcc tatcacctgc
     2881 gaaggaaggg cgtgccagcg ctgtacttgg gaccagacac ccccctcccc gatctcagag
     2941 cacttgcgag acggctcgga gccggagcgg tggttctgtc agctttgctt tccgagcctc
     3001 tcagggcgtt gccagacggc gcactgaaag acttggcacc tcgggtgttc ctgggagggc
     3061 aaggagccgg ccctgaagag gcccgacggc tcggggccga gtacatggaa gatctgaagg
     3121 gattggccga agcactgtgg cttccaagag gaccagagaa agaagcaatc ggaggtggag
     3181 gtagtgctag tggtggttct ggttccagat tagataaaag taaagtgatt aacagcgcat
     3241 tagagctgct taatgaggtc ggaatcgaag gtttaacaac ccgtaaactc gcccagaagc
     3301 taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa taagcgggct ttgctcgacg
     3361 ccttagccat tgagatgtta gataggcacc atactcactt ttgcccttta gaaggggaaa
     3421 gctggcaaga ttttttacgt aataacgcta aaagttttag atgtgcttta ctaagtcatc
     3481 gcgatggagc aaaagtacat ttaggtacac ggcctacaga aaaacagtat gaaactctcg
     3541 aaaatcaatt agccttttta tgccaacaag gtttttcact agagaatgca ttatatgcac
     3601 tcagcgctgt ggggcatttt actttaggtt gcgtattgga agatcaagag catcaagtcg
     3661 ctaaagaaga aagggaaaca cctactactg atagtatgcc gccattatta cgacaagcta
     3721 tcgaattatt tgatcaccaa ggtgcagagc cagccttctt attcggcctt gaattgatca
     3781 tatgcggatt agaaaaacaa cttaaatgtg aaagtgggtc cgctagtggt ggttctggtg
     3841 cccccccgac cgatgtcagc ctgggggacg agctccactt agacggcgag gacgtggcga
     3901 tggcgcatgc cgacgcgcta gacgatttcg atctggacat gttgggggac ggggattccc
     3961 cgggtccggg atttaccccc cacgactccg ccccctacgg cgctctggat atggccgact
     4021 tcgagtttga gcagatgttt accgatgccc ttggaattga cgagtacggt ggggctagtg
     4081 ctagcggatc caccggtgtc tagaaagctt tgaggccggc ctgcagggat cagcctcgac
     4141 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     4201 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     4261 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     4321 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     4381 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     4441 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4501 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4561 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4621 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4681 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4741 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4801 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4861 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4921 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4981 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     5041 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     5101 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     5161 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     5221 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     5281 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     5341 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5401 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5461 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5521 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5581 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5641 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5701 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5761 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5821 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5881 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5941 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     6001 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     6061 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     6121 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     6181 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     6241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6301 ctagttgtgg tttgtccaaa ctcatcaatg tatctta
//
