LOCUS       62056_3360        8224 bp DNA     circular SYN 01-SEP-2019
DEFINITION  Cloning vector PB-Gal4-Teton-cis, complete sequence.
ACCESSION   MK422200
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8224)
  AUTHORS   Chen B.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JAN-2019) Stem Cell Center, Institute of Blood 
REFERENCE   2  (bases 1 to 8224)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (17-JAN-2019) Stem Cell Center, Institute of Blood Transfusion, 
            Chinese Academy of Medical Sciences "
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1373..1391
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        1581..1619
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1684..2349
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             2419..2472
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     regulatory      2529..2653
                     /note="UAS; upstream activating sequences"
                     /regulatory_class="transcriptional_cis_regulatory_region"
     CDS             2970..3677
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    3678..3743
                     /note="P2A; 2A peptide"
     polyA_signal    3792..4016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4068..4397
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4445..4492
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4511..4882
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    5015..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6203..6219)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6227..6243)
                     /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(6251..6281)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6296..6317)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6605..7193)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7367..8224)
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggccact agacgagttt actccctatc
     1381 agtgatagag aacgatgtcg agtttactcc ctatcagtga tagagaacgt atgtcgagtt
     1441 tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac
     1501 gtatgtcgag tttatcccta tcagtgatag agaacgtatg tcgagtttac tccctatcag
     1561 tgatagagaa cgtatgtcga ggtaggcgtg tacggtggga ggcctatata agcagagctc
     1621 gtttagtgaa ccgtcagatc gcaccggtaa gctcgagcca ccatggagag cgacgagagc
     1681 ggcctgcccg ccatggagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc
     1741 gagctggtgg gcggcggaga gggcaccccc aagcagggcc gcatgaccaa caagatgaag
     1801 agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc
     1861 ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac
     1921 aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg
     1981 agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc
     2041 ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg
     2101 gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc
     2161 ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc
     2221 atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag
     2281 ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc
     2341 gccttcgcca gatcccgcgc tcagtcgtcc aattctgccg tggacggcac cgccggaccc
     2401 ggctccaccg gatctcgcga gggcagagga agtcttctaa catgcggtga cgtggaggag
     2461 aatcccggcc catttaaatc ctaaagaatt cggcgcgcca cgcgtccgag ctcttacgcg
     2521 ggtcgaagcg gagtactgtc ctccgagtgg agtactgtcc tccgagcgga gtactgtcct
     2581 ccgagtcgag ggtcgaagcg gagtactgtc ctccgagtgg agtactgtcc tccgagcgga
     2641 gtactgtcct ccgagtcgac tctagagggt atataatgga tctcgagata tcggagctcg
     2701 tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag
     2761 acaccgggac cgatccagcc tccgcggccg ggaacggtgc attggaacgc gcattccccg
     2821 tgttaattaa caggtaagtg tcttcctcct gtttccttcc cctgctattc tgctcaacct
     2881 tcctatcaga aactgcagta tctgtatttt tgctagcagt aatactaacg gttctttttt
     2941 tctcttcaca ggccaccaag cttgccacca tggtgagcaa gggcgaggag gataacatgg
     3001 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     3061 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     3121 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     3181 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     3241 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     3301 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     3361 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     3421 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     3481 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     3541 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     3601 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     3661 tggacgagct gtacaaggga tctggagcaa caaacttctc actactcaaa caagcaggtg
     3721 acgtggagga gaatcccggc ccgtttaaac aaacccacta gtgcggccgc aaacccgctg
     3781 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3841 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3901 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3961 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     4021 tgaggcggaa agaaccagct ggggctctag ggggtattct gtggaatgtg tgtcagttag
     4081 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4141 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4201 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4261 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4321 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4381 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     4441 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     4501 gaactaaacc atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc
     4561 cggagcggtc gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga
     4621 cttcgccggt gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt
     4681 ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga
     4741 gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat
     4801 cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca
     4861 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     4921 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4981 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     5041 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5101 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgggcca ttacggcccc
     5161 attggtatgg ctttttcccc gtatcccccc aggtgtctgc aggctcaaag agcagcgaga
     5221 agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt ccccgcacgc
     5281 tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg cggctcgctg
     5341 ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg gggggctgtc
     5401 cctgatatct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     5461 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     5521 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     5581 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     5641 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     5701 atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt tttccggctc
     5761 agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg tgccttttcc
     5821 cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg cattgacgtt
     5881 gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca cgcctttaac
     5941 ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt ccggacacag
     6001 ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga cagccggaac
     6061 tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta cgtcagcgag
     6121 gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga gttacccggc
     6181 gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6241 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6301 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6361 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6421 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6481 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6541 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6601 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6661 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6721 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6781 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6841 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6901 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6961 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     7021 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     7081 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7141 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7201 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7261 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7321 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7381 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7441 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7501 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7561 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7621 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7681 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7741 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7801 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7861 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7921 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7981 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     8041 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8101 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8161 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8221 tcat
//
