LOCUS       Exported                9964 bp ds-DNA     circular SYN 04-APR-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9964)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Aug 1, 2019 from SnapGene 4.3.8.1
            https://www.snapgene.com
COMMENT     Assemble of:
            Fragment  : pLD401 Orf2_His_Prescission-3xMyc3xFlag-Stop- SwaI in 
            pLD259 : BamHI at 5768 to PacI at 339.
            Fragment  : New DNA entry : PacI at 36 to BamHI at 79.
            Assembled on 2/24/2015
FEATURES             Location/Qualifiers
     source          1..9964
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..322
                     /label=Tet
     promoter        4..318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    83..101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    119..137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    155..173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    344..386
                     /label=MCS
     polyA_signal    388..522
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1273..3062
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     promoter        5814..5918
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5919..6779
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6950..7538
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        7833..8162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8013..8148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             8255..8854
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    9084..9218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    9338..9386
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
       61 tagagaacga tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttactc
      121 cctatcagtg atagagaacg tatgtcgagt ttactcccta tcagtgatag agaacgtatg
      181 tcgagtttat ccctatcagt gatagagaac gtatgtcgag tttactccct atcagtgata
      241 gagaacgtat gtcgaggtag gcgtgtacgg tgggaggcct atataagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagcggccgc gtttaaactt aattaaggcg cgccatgcgc
      361 acgtatacca cgtgcttaag ttcgaaggat ccagacatga taagatacat tgatgagttt
      421 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
      481 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
      541 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc
      601 tacaaatgtg gtatggctga ttatgatccg gctgcctcgc gcgtttcggt gatgacggtg
      661 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
      721 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
      781 tgaccggtcg actctagagc cccgccgccg gacgaactaa acctgactac ggcatctctg
      841 ccccttcttc gctggtacga ggagcgcttt tgttttgtat tcggggcagt gcatgtaatc
      901 ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg
      961 gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat
     1021 ttgccaacac tgagtggctt tcatcctgga gcagactttg catgctgtgg actgcaacac
     1081 aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt
     1141 acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg
     1201 tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc
     1261 ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta
     1321 accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag
     1381 taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta
     1441 catggggtca ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa
     1501 gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt
     1561 ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa
     1621 caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg
     1681 tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat
     1741 gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac
     1801 tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg
     1861 atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta
     1921 cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt
     1981 gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga
     2041 caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc
     2101 cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc
     2161 taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct
     2221 gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt
     2281 acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg
     2341 ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata
     2401 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     2461 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     2521 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     2581 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta
     2641 gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat
     2701 atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat
     2761 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     2821 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     2881 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     2941 atgctatcct catgcatata cagtcagcat atgataccca gtagtagagt gggagtgcta
     3001 tcctttgcat atgccgccac ctcccaaggg ggcgtgaatt ttcgctgctt gtccttttcc
     3061 tgcatgctgg ttgctcccat tcttaggtga atttaaggag gccaggctaa agccgtcgca
     3121 tgtctgattg ctcaccaggt aaatgtcgct aatgttttcc aacgcgagaa ggtgttgagc
     3181 gcggagctga gtgacgtgac aacatgggta tgcccaattg ccccatgttg ggaggacgaa
     3241 aatggtgaca agacagatgg ccagaaatac accaacagca cgcatgatgt ctactgggga
     3301 tttattcttt agtgcggggg aatacacggc ttttaatacg attgagggcg tctcctaaca
     3361 agttacatca ctcctgccct tcctcaccct catctccatc acctccttca tctccgtcat
     3421 ctccgtcatc accctccgcg gcagcccctt ccaccatagg tggaaaccag ggaggcaaat
     3481 ctactccatc gtcaaagctg cacacagtca ccctgatatt gcaggtagga gcgggctttg
     3541 tcataacaag gtccttaatc gcatccttca aaacctcagc aaatatatga gtttgtaaaa
     3601 agaccatgaa ataacagaca atggactccc ttagcgggcc aggttgtggg ccgggtccag
     3661 gggccattcc aaaggggaga cgactcaatg gtgtaagacg acattgtgga atagcaaggg
     3721 cagttcctcg ccttaggttg taaagggagg tcttactacc tccatatacg aacacaccgg
     3781 cgacccaagt tccttcgtcg gtagtccttt ctacgtgact cctagccagg agagctctta
     3841 aaccttctgc aatgttctca aatttcgggt tggaacctcc ttgaccacga tgctttccaa
     3901 accaccctcc ttttttgcgc ctgcctccat caccctgacc ccggggtcca gtgcttgggc
     3961 cttctcctgg gtcatctgcg gggccctgct ctatcgctcc cgggggcacg tcaggctcac
     4021 catctgggcc accttcttgg tggtattcaa aataatcggc ttcccctaca gggtggaaaa
     4081 atggccttct acctggaggg ggcctgcgcg gtggagaccc ggatgatgat gactgactac
     4141 tgggactcct gggcctcttt tctccacgtc cacgacctct ccccctggct ctttcacgac
     4201 ttccccccct ggctctttca cgtcctctac cccggcggcc tccactacct cctcgacccc
     4261 ggcctccact acctcctcga ccccggcctc cactgcctcc tcgaccccgg cctccacctc
     4321 ctgctcctgc ccctcctgct cctgcccctc ctcctgctcc tgcccctcct gcccctcctg
     4381 ctcctgcccc tcctgcccct cctgctcctg cccctcctgc ccctcctgct cctgcccctc
     4441 ctgcccctcc tcctgctcct gcccctcctg cccctcctcc tgctcctgcc cctcctgccc
     4501 ctcctgctcc tgcccctcct gcccctcctg ctcctgcccc tcctgcccct cctgctcctg
     4561 cccctcctgc tcctgcccct cctgctcctg cccctcctgc tcctgcccct cctgcccctc
     4621 ctgcccctcc tcctgctcct gcccctcctg ctcctgcccc tcctgcccct cctgcccctc
     4681 ctgctcctgc ccctcctcct gctcctgccc ctcctgcccc tcctgcccct cctcctgctc
     4741 ctgcccctcc tgcccctcct cctgctcctg cccctcctcc tgctcctgcc cctcctgccc
     4801 ctcctgcccc tcctcctgct cctgcccctc ctgcccctcc tcctgctcct gcccctcctc
     4861 ctgctcctgc ccctcctgcc cctcctgccc ctcctcctgc tcctgcccct cctcctgctc
     4921 ctgcccctcc tgcccctcct gcccctcctg cccctcctcc tgctcctgcc cctcctcctg
     4981 ctcctgcccc tcctgctcct gcccctcccg ctcctgctcc tgctcctgtt ccaccgtggg
     5041 tccctttgca gccaatgcaa cttggacgtt tttggggtct ccggacacca tctctatgtc
     5101 ttggccctga tcctgagccg cccggggctc ctggtcttcc gcctcctcgt cctcgtcctc
     5161 ttccccgtcc tcgtccatgg ttatcacccc ctcttctttg aggtccactg ccgccggagc
     5221 cttctggtcc agatgtgtct cccttctctc ctaggccatt tccaggtcct gtacctggcc
     5281 cctcgtcaga catgattcac actaaaagag atcaatagac atctttatta gacgacgctc
     5341 agtgaataca gggagtgcag actcctgccc cctccaacag cccccccacc ctcatcccct
     5401 tcatggtcgc tgtcagacag atccaggtct gaaaattccc catcctccga accatcctcg
     5461 tcctcatcac caattactcg cagcccggaa aactcccgct gaacatcctc aagatttgcg
     5521 tcctgagcct caagccaggc ctcaaattcc tcgtccccct ttttgctgga cggtagggat
     5581 ggggattctc gggacccctc ctcttcctct tcaaggtcac cagacagaga tgctactggg
     5641 gcaacggaag aaaagctggg tgcggcctgt gaggatcagc ttatcgatga taagctgtca
     5701 aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5761 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5821 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5881 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5941 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6001 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6061 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6121 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     6181 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6241 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6301 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6361 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6421 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt
     6481 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6541 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6601 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6661 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6721 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6781 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6841 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6901 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6961 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7021 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7081 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7141 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7201 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7261 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7321 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7381 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7441 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7501 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7561 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7621 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7681 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     7741 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     7801 ctgatgccgc atagttaagc cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     7861 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     7921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     7981 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     8041 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     8101 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     8161 aaaagcttgc atgcctgcag gtcggccgcc acgaccggtg ccgccaccat cccctgaccc
     8221 acgcccctga cccctcacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg
     8281 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     8341 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     8401 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     8461 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     8521 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     8581 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     8641 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     8701 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     8761 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     8821 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     8881 gaccgaaagg agcgcacgac cccatggctc cgaccgaagc cgacccgggc ggccccgccg
     8941 accccgcacc cgcccccgag gcccaccgac tctagaggat cataatcagc cataccacat
     9001 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     9061 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     9121 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     9181 tgtccaaact catcaatgta tcttatcatg tctggatcac tcgccgatag tggaaaccga
     9241 cgccccagca ctcgtccgag ggcaaaggaa taggggagat gggggaggct aactgaaaca
     9301 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     9361 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     9421 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     9481 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     9541 ccctgccata gccactggcc ccgtgggtta gggacggggt cccccatggg gaatggttta
     9601 tggttcgtgg gggttattat tttgggcgtt gcgtggggtc tggtccacga ctggactgag
     9661 cagacagacc catggttttt ggatggcctg ggcatggacc gcatgtactg gcgcgacacg
     9721 aacaccgggc gtctgtggct gccaaacacc cccgaccccc aaaaaccacc gcgcggattt
     9781 ctggcgtgcc aagctagtcg accaattctc atgtttgaca gcttatcatc gcagatccgg
     9841 gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat ctctagaagc
     9901 tgggtaccag ctgctagcaa gcttgctagc ggccgaggcg tatcacgagg ccctttcgtc
     9961 ttca
//
