LOCUS       62056_9375        7504 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 7504)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             733..756
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             757..2259
                     /codon_start=1
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     misc_feature    2280..2868
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    2934..2982
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3184..3612
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3626..3955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4022..4813
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4992..5125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5162..5178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5186..5202)
                     /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(5210..5240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5255..5276)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5564..6152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6326..7183)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7184..7288)
                     /label=AmpR promoter
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg
      721 cggccgctcg aggattacaa ggacgacgat gacaagatgc tgctgctgct gctgctgctg
      781 ggcctgaggc tacagctctc cctgggcatc atcccagttg aggaggagaa cccggacttc
      841 tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc tgcacagaca
      901 gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac ggtgacagct
      961 gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc cctggccatg
     1021 gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca tgtgccagac
     1081 agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca gaccattggc
     1141 ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga ggtcatctcc
     1201 gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac cacacgagtg
     1261 cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg gtactcggac
     1321 gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac gcagctcatc
     1381 tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt tcgcatggga
     1441 accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct ggacgggaag
     1501 aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact
     1561 gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct ctttgagcct
     1621 ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct gatggagatg
     1681 acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct cttcgtggag
     1741 ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact gactgagacg
     1801 atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga ggacacgctg
     1861 agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg
     1921 agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta cacggtcctc
     1981 ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga tgttaccgag
     2041 agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga cgaagagacc
     2101 cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc
     2161 gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct ggagccctac
     2221 accgcctgcg acctggcgcc ccccgccggc accaccgact gagggcccga tatctcgaca
     2281 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     2341 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     2401 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     2461 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     2521 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     2581 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     2641 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     2701 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     2761 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     2821 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg
     2881 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     2941 aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     3001 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     3061 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     3121 acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc
     3181 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat
     3361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3541 tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3601 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3961 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4021 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4081 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4141 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4201 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4261 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4321 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4381 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4441 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4501 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4561 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4621 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4681 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4741 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4801 tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca
     4861 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4921 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4981 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5041 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5101 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     5161 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     5221 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     5281 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     5341 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     5401 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5461 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5521 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5581 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5641 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5701 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     5761 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5821 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5881 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5941 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6001 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6061 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6121 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6181 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6241 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6301 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6361 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6421 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6481 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6541 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     6601 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     6661 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6721 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6781 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6841 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6901 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6961 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7021 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7081 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7141 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7201 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7261 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7321 acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc
     7381 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     7441 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     7501 tagg
//
