LOCUS       62056_401        6690 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 6690)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    46..94
                     /label=poly(A) signal
                     /note="synthetic polyadenylation signal"
     misc_feature    108..199
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from
                     the human alpha-2 globin gene"
     CDS             403..1119
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    1132..1707
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1716..3215
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="LLLLLLLGLRLQLSLGIILVEEENPDFWNREAAEALGAAKKLQPA
                     QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
                     VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
                     TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
                     MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLGERQGARYVWNRTELMQASLDPSVTHL
                     MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRHPRGFFLFVEGGRIDHGHHESRA
                     YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSFIGLAAGKAR
                     DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
                     GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    3779..3834
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4193..4209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4217..4233)
                     /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(4241..4271)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4286..4307)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        4365..4561
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     polyA_signal    4567..4701
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4940..5528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5702..6559)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6560..6664)
                     /label=AmpR promoter
ORIGIN
        1 gtcgagaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
       61 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
      121 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
      181 ggtgccagaa catttctcta tcgatagtcg accaattgct cgagttattg aggtcagctg
      241 aggtcaagtt acggatccgt aacttgacct cagaattctg aggtcaagtt acggatccgt
      301 aacttgacct cagaattcta tataatggaa gcttgaccgg agccgttagt tgcccgtcgg
      361 acgcgatttg cgaggcgctt tatgcgcgca ccggtcgcca ccatggtgag caagggcgag
      421 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      481 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      541 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      601 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      661 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      721 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      781 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      841 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      901 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      961 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1021 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1081 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggcca attctccctc
     1141 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1201 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1261 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1321 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1381 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1441 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1501 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     1561 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     1621 gtgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     1681 cctttgaaaa acacgatgat aatatggggt cgaggctgct gctgctgctg ctgctgggcc
     1741 tgaggctaca gctctccctg ggcatcatcc tagttgagga ggagaacccg gacttctgga
     1801 accgcgaggc agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg
     1861 ccaagaacct catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca
     1921 ggatcctaaa agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc
     1981 gcttcccata tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg
     2041 gagccacagc cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga
     2101 gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga
     2161 tgaatcgggc caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc
     2221 acgcctcgcc agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg
     2281 acgtgcctgc ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca
     2341 acatggacat tgatgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc
     2401 cagaccctga gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc
     2461 tggtgcagga atggctcggc gaacgccagg gtgcccggta cgtgtggaac cgcactgagc
     2521 tcatgcaggc ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag
     2581 acatgaaata cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag
     2641 aggctgccct gcgcctgctg agcagacacc cccgcggctt cttcctcttc gtggagggtg
     2701 gtcgcatcga ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca
     2761 tgttcgacga cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc
     2821 tcgtcactgc cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct
     2881 ccttcatcgg gctggccgct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat
     2941 acggaaacgg tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg
     3001 agagcgggag ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg
     3061 caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc
     3121 aggagcagac cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg
     3181 cctgcgacct ggcgcccccc gccggcacca ccgacgccgc gcacccgggg cggtccgtgg
     3241 tccccgcgtt gcttcctctg ctggccggga ccctgctgct gctggagacg gccactgctc
     3301 cctgagtgtc ccgtccctgg ggctcctgct tccccatccc ggagttctcc tgctccccac
     3361 ctcctgtcgt cctgcctggc ctccagcccg agtcgtcatc cccggagtcc ctatacagag
     3421 gtcctgccat ggaaccttcc cctccccgtg cgctctgggg actgagccca tgacaccaaa
     3481 cctgcccctt ggctgctctc ggactcccta ccccaacccc agggactgca ggttgtgccc
     3541 tgtggctgcc tgcaccccag gaaaggaggg ggctcaggcc atccagccac cacctacagc
     3601 ccagtgggta cggtcgagga tatcgcggcc ggccgcactc ctcaggtgca ggctgcctat
     3661 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3721 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3781 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3841 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3901 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta
     3961 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     4021 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4081 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4141 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     4201 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4261 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4321 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     4381 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4441 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4501 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4561 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4621 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4681 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4741 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4801 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4861 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5101 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5341 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5701 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5761 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5821 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5881 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5941 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6001 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6061 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6121 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6181 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6241 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6301 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6361 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6421 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6481 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6541 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6601 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6661 cgcgcacatt tccccgaaaa gtgccacctg
//
