LOCUS       Exported                8948 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    FUW-Sox10
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8948)
  AUTHORS   Guo W, Keckesova Z, Donaher JL, Shibue T, Tischler V, Reinhardt F, 
            Itzkovitz S, Noske A, Zurrer-Hardi U, Bell G, Tam WL, Mani SA, van 
            Oudenaarden A, Weinberg RA
  TITLE     Slug and Sox9 cooperatively determine the mammary stem cell state.
  JOURNAL   Cell. 2012 Mar 2;148(5):1015-28.
  PUBMED    22385965
REFERENCE   2  (bases 1 to 8948)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8948
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..271
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        304..342
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     339..363
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    471..495
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1897..1921)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1959..1979)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1964..2293
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2144..2279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2206..2225
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2303..2902
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2303..2322)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2799..2819
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2962..3550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3015..3035)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3433..3444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3553..3569)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3554..3570)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             4114..4294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(4320..4337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4326..4437
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4583..5171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4663..4682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5342..6202)
                     /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"
     primer_bind     5965..5984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6203..6307)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6382..6401)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6573..6952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6954..7152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7107..7127
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7170..7350
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7397..7522
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8015..8248
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8433..8477
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    8775..8892
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 tccctatcag tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag
       61 aaaagtgaaa gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
      121 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
      181 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
      241 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg gtacccgggt
      301 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
      361 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
      421 agcctccgcg gccccgaatt gcccagtgtg gtggaattct gcagatatca acaagtttgt
      481 acaaaaaagt tggcaccatg gcggaggagc aggatctatc ggaggtggag ctgagccccg
      541 tgggctcgga ggagccccgc tgcctgtccc cggggagcgc gccctcgcta gggcccgacg
      601 gcggcggcgg cggatcgggc ctgcgagcca gcccggggcc aggcgagctg ggcaaggtca
      661 agaaggagca gcaggacggc gaggcggacg atgacaagtt ccccgtgtgc atccgcgagg
      721 ccgtcagcca ggtgctcagc ggctacgact ggacgctggt gcccatgccc gtgcgcgtca
      781 acggcgccag caaaagcaag ccgcacgtca agcggcccat gaacgccttc atggtgtggg
      841 ctcaggcagc gcgcaggaag ctcgcggacc agtacccgca cctgcacaac gctgagctca
      901 gcaagacgct gggcaagctc tggaggctgc tgaacgaaag tgacaagcgc cccttcatcg
      961 aggaggctga gcggctccgt atgcagcaca agaaagacca cccggactac aagtaccagc
     1021 ccaggcggcg gaagaacggg aaggccgccc agggcgaggc ggagtgcccc ggtggggagg
     1081 ccgagcaagg tgggaccgcc gccatccagg cccactacaa gagcgcccac ttggaccacc
     1141 ggcacccagg agagggctcc cccatgtcag atgggaaccc cgagcacccc tcaggccaga
     1201 gccatggccc acccacccct ccaaccaccc cgaagacaga gctgcagtcg ggcaaggcag
     1261 acccgaagcg ggacgggcgc tccatggggg agggcgggaa gcctcacatc gacttcggca
     1321 acgtggacat tggtgagatc agccacgagg taatgtccaa catggagacc tttgatgtgg
     1381 ctgagttgga ccagtacctg ccgcccaatg ggcacccagg ccatgtgagc agctactcag
     1441 cagccggcta tgggctgggc agtgccctgg ccgtggccag tggacactcc gcctggatct
     1501 ccaagccacc aggcgtggct ctgcccacgg tctcaccacc tggtgtggat gccaaagccc
     1561 aggtgaagac agagaccgcg gggccccagg ggcccccaca ctacaccgac cagccatcca
     1621 cctcacagat cgcctacacc tccctcagcc tgccccacta tggctcagcc ttcccctcca
     1681 tctcccgccc ccagtttgac tactctgacc atcagccctc aggaccctat tatggccact
     1741 cgggccaggc ctctggcctc tactcggcct tctcctatat ggggccctcg cagcggcccc
     1801 tctacacggc catctctgac cccagcccct cagggcccca gtcccacagc cccacacact
     1861 gggagcagcc agtatatacg acactgtccc ggccctaacc aactttcttg tacaaagtgg
     1921 ttgatatcca gcacagtggt cgacgataaa tgatccgtgg aatgtgtgtc agttagggtg
     1981 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2041 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2101 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2161 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2221 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2281 aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2341 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     2401 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     2461 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     2521 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     2581 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     2641 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     2701 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     2761 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     2821 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     2881 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     2941 cgcacgaccc catgcatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3001 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3061 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3121 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3181 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3241 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3301 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3361 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3421 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3481 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3541 gcctccccgc atcgataccg tcgacctcga gacctagaaa aacatggagc aatcacaagt
     3601 agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag
     3661 gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta
     3721 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     3781 agacaagata tccttgatct gtggatctac cacacacaag gctacttccc tgattggcag
     3841 aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg ctacaagcta
     3901 gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac ccgcttgtta
     3961 caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga gtggaggttt
     4021 gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggaata acttcgtata
     4081 ryrtayryta tacgaagtta tccggactgt actgggtctc tctggttaga ccagatctga
     4141 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     4201 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     4261 agaccctttt agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc
     4321 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     4381 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     4441 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     4501 ggattgggaa gacaatagca ggcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     6061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     6121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     6181 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     6241 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     6301 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     6361 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6421 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6481 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6541 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     6601 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     6661 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     6721 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     6781 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6841 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6901 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     6961 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     7021 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     7081 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     7141 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
     7201 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     7261 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     7321 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
     7381 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
     7441 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
     7501 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
     7561 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     7621 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     7681 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     7741 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     7801 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     7861 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     7921 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     7981 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     8041 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     8101 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     8161 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     8221 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     8281 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     8341 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     8401 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     8461 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     8521 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     8581 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     8641 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     8701 gatccattcg attagtgaac ggatcggcac tgcgtgcgcc aattctgcag acaaatggca
     8761 gtattcatcc acaattttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga
     8821 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa
     8881 attcaaaatt ttcgggttta ttacagggac agcagagatc cagtttggtt agatctcgag
     8941 tttaccac
//
