LOCUS       V030261                 9914 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030261
VERSION     V030261
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     polyA_signal    216..350
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        398..502
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             503..1363
                     /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"
     rep_origin      1534..2119
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     protein_bind    2407..2428
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     promoter        2443..2473
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    2481..2497
                     /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"
     primer_bind     2505..2521
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     LTR             2650..3283
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    3490..4078
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     CDS             complement(4092..4691)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     promoter        complement(4700..5029)
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             complement(5040..5786)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of
                     doxycycline"
                     /transl_table=1
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG*"
                     /label="Tet-On® 3G"
     promoter        complement(5805..6315)
                     /note="human phosphoglycerate kinase 1 promoter"
                     /label="hPGK promoter"
     promoter        6332..6699
                     /note="3rd-generation Tet-responsive promoter that can be
                     activated by binding of Tet-On® 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription
                     factors"
                     /label="TRE3GS promoter"
     CDS             6712..7410
                     /label="BCL2L1(NM_138578)"
                     /note="BCL2L1(NM_138578)"
                     /gene="BCL2L1"
     polyA_signal    7594..7728
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     misc_feature    7772..7887
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     CDS             complement(8150..8194)
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    8379..8612
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     misc_feature    9109..9234
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     LTR             9281..9914
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
ORIGIN
        1 actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaaact actaagtttg
       61 tagtacatat ttaacaaata caatttcttt aaaatgaaaa taattcagag gaatcacagg
      121 tttagagtaa atgaaaccac taggtaattg gcagtggtaa tagggtatgg ggtgggaagt
      181 ttgggatgat tttggttagc ttgagttatc cagttgatcc agacatgata agatacattg
      241 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      301 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt gatctcccga
      361 tccgtcgacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      421 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      481 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      541 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      601 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      661 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      721 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      781 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      841 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      901 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      961 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1021 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1081 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1141 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1201 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1261 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1321 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1381 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1441 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1501 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1561 ctgctgcttg caaacaaaaa aaccgctacc agcggtggtt tgtttgccgg atcaagagct
     1621 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1681 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1741 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1801 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     1861 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     1921 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     1981 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2041 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2101 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2161 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2221 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2281 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2341 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     2401 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     2461 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     2521 ccatgattac gccaagctct agctagaggt cgacggtaaa acgctagcaa tgtcaaggcc
     2581 tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata agatgacatg
     2641 aactactact gctagagatt ttccacactg actaaaaggg tctgagggat ctctagttac
     2701 cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag ctttattgag
     2761 gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc tggtctaacc
     2821 agagagaccc agtacaggca aaaagcagct gcttatatgc aggatctgag ggctcgccac
     2881 tccccagtcc cgcccaggcc acgcctccct ggaaagtccc cagcggaaag tcccttgtag
     2941 caagctcgat atcagcagtt cttgaagtac tccggatgca gctctcgggc cacgtgatga
     3001 aatgctaggc ggctgtcaaa cctccactct aacacttctc tctccgggtc atccatccca
     3061 tgcaggctca cagggtgtaa caagctggtg ttctctcctt tattggcctc ttctacctta
     3121 tctggctcaa ctggtactag cttgtagcac catccaaagg tcagtggata tctgacccct
     3181 ggccctggtg tgtagttctg ctaatcaggg aagtagcctt gtgtgtggta gatccacaga
     3241 tcaaggatat cttgtcttcg ttgggagtga attagccctt ccagtcccct cttttctttt
     3301 aaaaagtggc taagatctac agctgccttg taagtcattg gtcttaaagg tacctgaggt
     3361 gtgactggaa aacccacctc ctcctcctct tgtgcttcta gccaggcaca atcagcattg
     3421 gtagctgctg tattgctact tgtgattgct ccatgttttt ctaggtctcg actgcagaat
     3481 taattccagg cggggaggcg gcccaaaggg agatccgact cgtctgaggg cgaaggcgaa
     3541 gacgcggaag aggccgcaga gccggcagca ggccgcggga aggaaggtcc gctggattga
     3601 gggccgaagg gacgtagcag aaggacgtcc cgcgcagaat ccaggtggca acacaggcga
     3661 gcagccatgg aaaggacgtc agcttccccg acaacaccac ggaattgtca gtgcccaaca
     3721 gccgagcccc tgtccagcag cgggcaaggc aggcggcgat gagttccgcc gtggcaatag
     3781 ggagggggaa agcgaaagtc ccggaaagga gctgacaggt ggtggcaatg ccccaaccag
     3841 tgggggttgc gtcagcaaac acagtgcaca ccacgccacg ttgcctgaca acgggccaca
     3901 actcctcata aagagacagc aaccaggatt tatacaagga ggagaaaatg aaagccatac
     3961 gggaagcaat agcatgatac aaaggcatta aagcagcgta tccacatagc gtaaaaggag
     4021 caacatagtt aagaatacca gtcaatcttt cacaaatttt gtaatccaga ggttgattgt
     4081 tccagacgcg ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     4141 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     4201 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     4261 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg
     4321 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     4381 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     4441 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     4501 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     4561 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     4621 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     4681 tactcggtca tggtcgcgtt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     4741 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     4801 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     4861 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     4921 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     4981 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttcgcgcgtt
     5041 tacccgggga gcatgtcaag gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg
     5101 tcaaagtcgt caagggcatc ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg
     5161 gtcggcccgc cgctttcgca ctttagctgt ttctccaggc cacatatgat tagttccagg
     5221 ccgaaaagga aggcaggttc ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga
     5281 agtgggggca tagaatcggt ggtaggtgtc tctctttcct cttttgctac ttgatgctcc
     5341 tgttcctcca atacgcagcc cagtgtaaag tggcccacgg cggacagagc gtacagtgcg
     5401 ttctccaggg agaagccttg ctgacacagg aacgcgagct gattttccag ggtttcgtac
     5461 tgtttctctg ttgggcgggt gccgagatgc actttagccc cgtcgcgatg tgagaggaga
     5521 gcacagcggt atgacttggc gttgttccgc agaaagtctt gccatgactc gccttccagg
     5581 gggcaggagt gggtatgatg cctgtccagc atctcgattg gcagggcatc gagcagggcc
     5641 cgcttgttct tcacgtgcca gtacagggta ggctgctcaa ctcccagctt ttgagcgagt
     5701 ttccttgtcg tcaggccttc gataccgact ccattgagta attccagagc agagtttatg
     5761 actttgctct tgtccagtct agacatggta attcgatgat ccccctgggg agagaggtcg
     5821 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg
     5881 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc
     5941 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc
     6001 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg
     6061 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga
     6121 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg
     6181 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc
     6241 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg
     6301 cgcaacccca accccgtgga attatcacct cgagtttact ccctatcagt gatagagaac
     6361 gtatgaagag tttactccct atcagtgata gagaacgtat gcagacttta ctccctatca
     6421 gtgatagaga acgtataagg agtttactcc ctatcagtga tagagaacgt atgaccagtt
     6481 tactccctat cagtgataga gaacgtatct acagtttact ccctatcagt gatagagaac
     6541 gtatatccag tttactccct atcagtgata gagaacgtat aagctttgct tatgtaaacc
     6601 agggcgccta taaaagagtg ctgatttttt gagtaaactt caattccaca acacttttgt
     6661 cttataccaa ctttccgtac cacttcctac cctcgtaaag aattcgccac catgtctcag
     6721 agcaaccggg agctggtggt tgactttctc tcctacaagc tttcccagaa aggatacagc
     6781 tggagtcagt ttagtgatgt ggaagagaac aggactgagg ccccagaagg gactgaatcg
     6841 gagatggaga cccccagtgc catcaatggc aacccatcct ggcacctggc agacagcccc
     6901 gcggtgaatg gagccactgg ccacagcagc agtttggatg cccgggaggt gatccccatg
     6961 gcagcagtaa agcaagcgct gagggaggca ggcgacgagt ttgaactgcg gtaccggcgg
     7021 gcattcagtg acctgacatc ccagctccac atcaccccag ggacagcata tcagagcttt
     7081 gaacaggtag tgaatgaact cttccgggat ggggtaaact ggggtcgcat tgtggccttt
     7141 ttctccttcg gcggggcact gtgcgtggaa agcgtagaca aggagatgca ggtattggtg
     7201 agtcggatcg cagcttggat ggccacttac ctgaatgacc acctagagcc ttggatccag
     7261 gagaacggcg gctgggatac ttttgtggaa ctctatggga acaatgcagc agccgagagc
     7321 cgaaagggcc aggaacgctt caaccgctgg ttcctgacgg gcatgactgt ggccggcgtg
     7381 gttctgctgg gctcactctt cagtcggaaa tgaggatcca gaccacctcc cctgcgagct
     7441 aagctggaca gccaatgacg ggtaagagag tgacattttt cactaaccta agacaggagg
     7501 gccgtcagag ctactgccta atccaaagac gggtaaaagt gataaaaatg tatcactcca
     7561 acctaagaca ggcgcagctt ccgagggatt tgagatccag acatgataag atacattgat
     7621 gagtttggac aaaccaaaac tagaatgcag tgaaaaaaat gccttatttg tgaaatttgt
     7681 gatgctattg ccttatttgt aaccattata agctgcaata aacaagttaa gtcgataaac
     7741 tggatctctg ctgtccctgt aataaacccg aaaattttga atttttgtaa tttgtttttg
     7801 tagttcttta gtttgtatgt ctgttgctat tatgtctact attctttccc ctgcactgta
     7861 ccccccaatc cccccttttc ttttaaaggc gataccgtcg agatccgttc actaatcgaa
     7921 tggatctgtc tctgtctctc tctccacctt cttcttctat tccttcgggc ctgtcgggtc
     7981 ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc ctaactctat
     8041 tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct actatcatta
     8101 tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac aaacttgccc
     8161 atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc gattcttcaa
     8221 ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc atccaggtcg
     8281 tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa ggcacagcag
     8341 tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga tcctttaggt
     8401 atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact gtgagttgca
     8461 acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc actataccag
     8521 acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata gtgcttcctg
     8581 ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt tctctctgca
     8641 ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt actactttat
     8701 atttatataa ttcacttctc caattgtccc tcatatctcc tcctccaggt ctgaagatca
     8761 gcggccggcc gcttgctgtg cggtggtctt acttttgttt tgctcttcct ctatcttgtc
     8821 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
     8881 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
     8941 cagtatttgt ctacagcctt ctgatgtttc taacaggcca ggattaactg cgaatcgttc
     9001 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
     9061 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
     9121 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
     9181 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
     9241 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
     9301 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
     9361 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
     9421 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaaagcagc
     9481 tgcttatatg caggatctga gggctcgcca ctccccagtc ccgcccaggc cacgcctccc
     9541 tggaaagtcc ccagcggaaa gtcccttgta gcaagctcga tatcagcagt tcttgaagta
     9601 ctccggatgc agctctcggg ccacgtgatg aaatgctagg cggctgtcaa acctccactc
     9661 taacacttct ctctccgggt catccatccc atgcaggctc acagggtgta acaagctggt
     9721 gttctctcct ttattggcct cttctacctt atctggctca actggtacta gcttgtagca
     9781 ccatccaaag gtcagtggat atctgacccc tggccctggt gtgtagttct gctaatcagg
     9841 gaagtagcct tgtgtgtggt agatccacag atcaaggata tcttgtcttc tttgggagtg
     9901 aattagccct tcca
//
