LOCUS       62056_19605        8861 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8861)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             397..988
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    1051..1408
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1482..1889
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="VTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTFNVG
                     WPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPP
                     LPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    1899..2269
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     protein_bind    2286..2556
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2589..2627
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2714..3430
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3449..4034
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4035..5039
                     /codon_start=1
                     /label=tTA
                     /note="tetracycline-controlled transactivator, comprising a
                     fusion of the tetracycline repressor TetR with the 
                     C-terminal activation domain of herpes simplex virus VP16"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTI
                     EGLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHA
                     DALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(6232..6248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6723..6827
                     /label=AmpR promoter
     CDS             6828..7685
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7859..8447
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8735..8756
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8771..8801
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8809..8825
                     /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)."
     primer_bind     8833..8849
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctttgct cttaggagtt tcctaataca tcccaaactc aaatatataa agcatttgac
       61 ttgttctatg ccctaggggg cggggggaag ctaagccagc tttttttaac atttaaaatg
      121 ttaattccat tttaaatgca cagatgtttt tatttcataa gggtttcaat gtgcatgaat
      181 gctgcaatat tcctgttacc aaagctagta taaataaaaa tagataaacg tggaaattac
      241 ttagagtttc tgtcattaac gtttccttcc tcagttgaca acataaatgc gctgctgagc
      301 aagccagttt gcatctgtca ggatcaattt cccattatgc cagtcatatt aattactagt
      361 caattagttg atttttattt ttgacatata catgtgaatg aaagacccca cctgtaggtt
      421 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
      481 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
      541 atctgtggta agcagttcct gccccgctca gggccaagaa cagatggaac agctgaatat
      601 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg ctcagggcca agaacagatg
      661 gtccccagat gcggtccagc cctcagcagt ttctagagaa ccatcagatg tttccagggt
      721 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
      781 gcttctgttc gcgcgcttat gctccccgag ctcaataaaa gagcccacaa cccctcactc
      841 ggggcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc
      901 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
      961 gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg agacccctgc
     1021 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
     1081 cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt tagctaacta
     1141 gctctgtatc tggcggaccc gtggtggaac tgacgagttc ggaacacccg gccgcaaccc
     1201 tgggagacgt cccagggact tcgggggccg tttttgtggc ccgacctgag tcctaaaatc
     1261 ccgatcgttt aggactcttt ggtgcacccc ccttagagga gggatatgtg gttctggtag
     1321 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttgggacc
     1381 gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc tgtctgactg
     1441 tgtttctgta tttgtctgaa aatatgggcc cgggctagcc tgttaccact cccttaagtt
     1501 tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca
     1561 agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc
     1621 cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac
     1681 ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt
     1741 ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc
     1801 catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt
     1861 atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc ttatatgggg
     1921 cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact aacagcccct
     1981 ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg agacctctgg
     2041 cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac cgagtcggcg
     2101 acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg aaaggacctt
     2161 acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca gcttggatac
     2221 acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctagc cctcgagttt
     2281 accactccct atcagtgata gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga
     2341 tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc
     2401 gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat
     2461 cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg
     2521 aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gctcggtacc
     2581 cgggtcgagg taggcgtgta cggtgggagg cctatataag cagagctcgt ttagtgaacc
     2641 gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
     2701 gatccgacct gccatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2761 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     2821 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     2881 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     2941 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3001 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3061 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3121 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     3181 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     3241 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     3301 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     3361 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     3421 gctgtacaag taaagcggcc gcggatcccc cctctccctc ccccccccct aacgttactg
     3481 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     3541 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3601 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3661 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3721 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3781 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3841 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3901 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     3961 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     4021 agcttgccac aaccatgtct agattagata aaagtaaagt gattaacagc gcattagagc
     4081 tgcttaatga ggtcggaatc gaaggtttaa caacccgtaa actcgcccag aagctaggtg
     4141 tagagcagcc tacattgtat tggcatgtaa aaaataagcg ggctttgctc gacgccttag
     4201 ccattgagat gttagatagg caccatactc acttttgccc tttagaaggg gaaagctggc
     4261 aagatttttt acgtaataac gctaaaagtt ttagatgtgc tttactaagt catcgcgatg
     4321 gagcaaaagt acatttaggt acacggccta cagaaaaaca gtatgaaact ctcgaaaatc
     4381 aattagcctt tttatgccaa caaggttttt cactagagaa tgcattatat gcactcagcg
     4441 ctgtggggca ttttacttta ggttgcgtat tggaagatca agagcatcaa gtcgctaaag
     4501 aagaaaggga aacacctact actgatagta tgccgccatt attacgacaa gctatcgaat
     4561 tatttgatca ccaaggtgca gagccagcct tcttattcgg ccttgaattg atcatatgcg
     4621 gattagaaaa acaacttaaa tgtgaaagtg ggtccgcgta cagccgcgcg cgtacgaaaa
     4681 acaattacgg gtctaccatc gagggcctgc tcgatctccc ggacgacgac gcccccgaag
     4741 aggcggggct ggcggctccg cgcctgtcct ttctccccgc gggacacacg cgcagactgt
     4801 cgacggcccc cccgaccgat gtcagcctgg gggacgagct ccacttagac ggcgaggacg
     4861 tggcgatggc gcatgccgac gcgctagacg atttcgatct ggacatgttg ggggacgggg
     4921 attccccggg tccgggattt accccccacg actccgcccc ctacggcgct ctggatatgg
     4981 ccgacttcga gtttgagcag atgtttaccg atgcccttgg aattgacgag tacggtgggt
     5041 agggggcgcg aagatccgga ttagtccaat ttgttaaaga caggatatca gtggtccagg
     5101 ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa
     5161 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     5221 ttggcaagct agctcgaccc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     5281 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     5341 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     5401 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5461 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5521 cgggggtctt tcacacatgc agcatgtatc aaaattaatt tggttttttt tcttaagtat
     5581 ttacattaaa tggccatagt acttaaagtt acattggctt ccttgaaata aacatggagt
     5641 attcagaatg tgtcataaat atttctaatt ttaagatagt atctccattg gctttctact
     5701 ttttctttta tttttttttg tcctctgtct tccatttgtt gttgttgttg tttgtttgtt
     5761 tgtttgttgg ttggttggtt aatttttttt taaagatcct acactatagt tcaagctaga
     5821 ctattagcta ctctgtaacc cagggtgacc ttgaagtcat gggtagcctg ctgttttagc
     5881 cttcccacat ctaagattac aggtatgagc tatcattttt ggtatattga ttgattgatt
     5941 gattgatgtg tgtgtgtgtg attgtgtttg tgtgtgtgac tgtgaaaatg tgtgtatggg
     6001 tgtgtgtgaa tgtgtgtatg tatgtgtgtg tgtgagtgtg tgtgtgtgtg tgtgcatgtg
     6061 tgtgtgtgtg actgtgtcta tgtgtatgac tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg
     6121 tgtgtgtgtg tgtgtgttgt gaaaaaatat tctatggtag tgagagccaa cgctccggct
     6181 caggtgtcag gttggttttt gagacagagt ctttcactta gcttggaatt cactggccgt
     6241 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     6301 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     6361 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     6421 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     6481 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     6541 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     6601 ttcaccgtca tcaccgaaac gcgcgatgac gaaagggcct cgtgatacgc ctatttttat
     6661 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     7021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     7081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7981 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     8041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     8101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8521 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8581 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8641 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8701 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8761 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     8821 aacaatttca cacaggaaac agctatgacc atgattacgc c
//
