LOCUS       62056_8525        5746 bp DNA     circular SYN 07-JUL-2019
DEFINITION  Cloning vector pDN-MMa6h, complete sequence.
ACCESSION   MK816964
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5746)
  AUTHORS   Farquhar KS, Charlebois DA, Szenk M, Cohen J, Nevozhay D, Balazsi G.
  TITLE     Role of network-mediated stochasticity in mammalian drug resistance
  JOURNAL   Nat Commun 10 (1), 2766 (2019)
  PUBMED    31235692
REFERENCE   2  (bases 1 to 5746)
  AUTHORS   Farquhar KS, Charlebois DA, Szenk M, Cohen J, Nevozhay D, Balazsi G.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2019) Laufer Center for Physical and Quantitative 
            Biology, Stony Brook University, 100 Nicolls Road, Stony Brook, NY 
            11790, USA
REFERENCE   3  (bases 1 to 5746)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "2766"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-APR-2019) Laufer Center for Physical and Quantitative Biology, 
            Stony Brook University, 100 Nicolls Road, Stony Brook, NY 11790, 
            USA"
FEATURES             Location/Qualifiers
     source          1..5746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             921..1664
                     /codon_start=1
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    1711..1935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1981..2409
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2423..2752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        2800..2847
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2866..3261
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3422..3555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3592..3608)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3616..3632)
                     /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(3640..3670)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3685..3706)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3994..4579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4753..5610)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5611..5715)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      841 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      901 tccgcggccc cgaattcacc atgtctagac tggacaagag caaagtcata aacggcgctc
      961 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     1021 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     1081 ccctgccaat cgagatgctg gacaggcatc atacccactt ctgccccctg gaaggcgagt
     1141 catggcaaga ctttctgcgg aacaacgcca agtcattccg ctgtgctctc ctctcacatc
     1201 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     1261 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     1321 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     1381 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg agacaagcaa
     1441 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     1501 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccggccgac gcccttgacg
     1561 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     1621 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaactc gagtctagag
     1681 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1741 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1801 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1861 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1921 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     1981 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2041 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2101 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2161 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2221 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2281 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2341 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2401 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2461 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2521 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2581 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2641 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2701 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     2761 gggagcttgt atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca
     2821 tagtatatcg gcatagtata atacgacaag gtgaggaact aaaccatggc caagcctttg
     2881 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     2941 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     3001 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     3061 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     3121 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     3181 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     3241 tctggttatg tgtgggaggg ctaagcactt cgtggccgag gagcaggact gacacgtgct
     3301 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3361 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3421 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3481 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3541 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3601 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3661 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3721 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3781 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3841 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3901 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3961 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4021 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4081 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4141 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4201 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4261 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4321 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4381 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4441 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4501 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     4561 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4621 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4681 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     4741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5341 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     5401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     5521 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     5581 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     5641 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     5701 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
