LOCUS       V037581                 8125 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037581
VERSION     V037581
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3615
                     /label="Bcl11b(NM_001079883)"
                     /note="Bcl11b(NM_001079883)"
                     /gene="Bcl11b"
     CDS             3622..3687
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3725..3949
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3995..4423
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4437..4766
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4833..5627
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5801..5922
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5971..5987)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5995..6011
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6019..6049)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6064..6085
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6373..6958)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7129..7989)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7990..8094)
                     /gene="bla"
                     /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 gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgtccc gccgcaaaca gggcaacccg cagcacttgt cccagaggga actcatcacg
     1021 ccagaggctg accatgtgga ggctaccatc ctcgaggaag acgagggtct ggagatagag
     1081 gagcctagca gcctggggct gatggtggga ggccccgacc ctgatctact cacctgtggc
     1141 cagtgtcaga tgaacttccc gctgggggac atcctggttt ttatagagca caagaagaaa
     1201 cagtgtggag gcctgggccc ctgctacgac aaggtcctgg acaagagcag tccacctccc
     1261 tcctctcgct ctgagctcag gagagtatct gagccagtgg agatcgggat ccaggtcacc
     1321 cctgatgaag atgaccacct actgtcaccc acgaaaggca tctgtcccaa gcaggagaac
     1381 attgcagggc cgtgcaggcc tgcccagctg ccatcgatgg cccccatagc tgcctcctct
     1441 tcccaccctc ccacctccgt gattacttca cctctgcgtg ccctgggcgt tctcccaccc
     1501 tgtttcccgc tgccttgctg tggtgcacgc cccatctcgg gcgacgggac tcagggtgag
     1561 ggtcagatgg aggctccctt tggatgccag tgtgagttgt caggtaaaga tgagccttcc
     1621 agctacattt gcacaacatg caagcagccc ttcaacagcg cctggttcct gctgcagcac
     1681 gcacagaaca cacatggctt ccgaatctac ctggagcctg ggccggccag cacctcgctc
     1741 acgcccaggc tcaccatccc gccaccgctc gggccggaga ccgtggcgca gtccccactc
     1801 atgaatttcc tgggggacag caatcctttc aacctgctgc gcatgacggg ccccatcctg
     1861 cgggaccacc ctggcttcgg tgagggccgc ttgccaggta cgccaccgct cttcagccca
     1921 ccgccacgcc atcacttgga cccacaccgc ctcagtgcag aggagatggg gctcgtggcc
     1981 cagcacccca gtgccttcga ccgagtcatg cgcctgaacc ccatggccat agactctcct
     2041 gccatggact tctcccggcg gctgcgagaa ctggccggca acagctccac gccgccgccc
     2101 gtgtccccag gccgtggcaa ccctatgcac cggctgctga accctttcca gcccagtccc
     2161 aagtccccgt tcctcagcac gccaccgctg ccacccatgc ctgcgggcac accgccaccg
     2221 cagccgcctg ccaagagcaa gtcctgtgag ttctgcggca agaccttcaa gttccagagc
     2281 aatctcatcg tgcaccggcg cagccacacg ggcgagaagc cctacaagtg ccagctgtgc
     2341 gaccatgcgt gctcgcaggc gagcaagctc aagcgccaca tgaagacgca catgcacaag
     2401 gcgggctctc tggctggccg ctcagacgac gggctctcag ctgccagctc ccctgagccg
     2461 ggcaccagcg agctgccagg tgacctgaaa gcggccgatg gcgacttccg ccaccatgag
     2521 agcgacccat ctctgggccc cgagcctgag gacgacgagg acgaggagga ggaagaagag
     2581 gagctgctgc tggagaacga gagccggcct gagtcgagct tcagcatgga ctcggagctg
     2641 ggccgtggcc gcgagaacgg aggtggcgtg ccaccggggg tggcgggcgc aggggctgca
     2701 gctgcggctc tggcggatga gaaggctctg gccctgggca aggtgatgga ggacgcaggg
     2761 ctgggcgcac tgccgcagta tggggagaag cggggcgcct tcctgaagcg tgcaggcgac
     2821 acgggtgatg ccggagctgt tggctgtggg gacgcgggtg caccgggtgc agtgaacggg
     2881 cgcggcgggg ccttcgcgcc aggcgcagag ccctttccag ctctcttccc acgcaagcca
     2941 gcaccgctgc ccagccctgg gctcggtggt cccgcgctgc acgcggccaa gcgcatcaag
     3001 gtggagaaag acctggagct gccacctgcc gccctcatcc catctgagaa cgtgtactcg
     3061 cagtggctcg tgggctacgc agcatcgcgc cacttcatga aggacccatt cctgggcttc
     3121 acggatgcgc gccagtcgcc tttcgccaca tcgtcggaac attcctctga gaacggcagc
     3181 ctgcgcttct caacgccacc cggggacctg ctggacggcg ggctgtccgg gcgcagtggc
     3241 acggcgagcg ggggcagcac acctcacctg ggtggtccgg gtcctgggag gccgagctcc
     3301 aaggagggcc gccgcagcga cacatgtgag tactgcggca aggtcttcaa gaactgtagc
     3361 aacctgacgg tgcaccggag gagccacacc ggcgagcggc cttacaagtg cgagctgtgc
     3421 aactacgcgt gcgcgcagag cagcaagctc acgcgccaca tgaagacgca cgggcagatc
     3481 ggcaaggagg tgtaccgctg cgacatctgc cagatgccct tcagcgtcta cagcaccctg
     3541 gagaaacaca tgaaaaagtg gcacggtgaa cacttgctga ctaatgatgt caaaatcgag
     3601 caggctgaga ggagcctcga ggactacaaa gaccatgacg gtgattataa agatcatgac
     3661 atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg ctgatcagcc
     3721 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3781 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3841 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3901 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3961 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4141 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4441 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4561 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4621 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4681 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4741 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4801 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4861 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4921 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4981 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5041 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5101 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5161 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5221 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5281 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5341 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5401 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5461 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5521 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5581 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5641 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5701 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5761 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5821 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5881 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5941 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     6001 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     6061 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     6121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     6181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     6241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     6301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     6361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     6421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6901 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8101 tccccgaaaa gtgccacctg acgtc
//
