LOCUS Exported 5583 bp ds-DNA circular SYN 06-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..5583
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 94..103
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 100..1749
/codon_start=1
/product="firefly luciferase"
/label=luciferase
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
CDS 1756..1803
/codon_start=1
/product="non-ORF yeast peptide conferring
ubiquitin-dependent degradation"
/label=hCL1
/note="human codon-optimized"
/translation="ACKNWFSSLSHFVIHL"
CDS 1807..1926
/codon_start=1
/product="PEST degradation sequence from mouse ornithine
decarboxylase"
/label=hPEST
/note="human codon-optimized"
/translation="SHGFPPEVEEQAAGTLPMSCAQESGMDRHPAACASARINV"
polyA_signal 1975..2096
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 2290..2647
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2498..2633
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2678..3277
/codon_start=1
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 3302..3350
/note="synthetic polyadenylation signal"
rep_origin complement(3677..4265)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4465..5325)
/codon_start=1
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
polyA_signal 5430..5478
/note="synthetic polyadenylation signal"
misc_feature 5492..5583
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
ORIGIN
1 ggcctaactg gccggtacct gagctcgcta gcctcgagga tatcaagatc tggcctcggc
61 ggccaagctt ggcaatccgg tactgttggt aaagccacca tggaagatgc caaaaacatt
121 aagaagggcc cagcgccatt ctacccactc gaagacggga ccgccggcga gcagctgcac
181 aaagccatga agcgctacgc cctggtgccc ggcaccatcg cctttaccga cgcacatatc
241 gaggtggaca ttacctacgc cgagtacttc gagatgagcg ttcggctggc agaagctatg
301 aagcgctatg ggctgaatac aaaccatcgg atcgtggtgt gcagcgagaa tagcttgcag
361 ttcttcatgc ccgtgttggg tgccctgttc atcggtgtgg ctgtggcccc agctaacgac
421 atctacaacg agcgcgagct gctgaacagc atgggcatca gccagcccac cgtcgtattc
481 gtgagcaaga aagggctgca aaagatcctc aacgtgcaaa agaagctacc gatcatacaa
541 aagatcatca tcatggatag caagaccgac taccagggct tccaaagcat gtacaccttc
601 gtgacttccc atttgccacc cggcttcaac gagtacgact tcgtgcccga gagcttcgac
661 cgggacaaaa ccatcgccct gatcatgaac agtagtggca gtaccggatt gcccaagggc
721 gtagccctac cgcaccgcac cgcttgtgtc cgattcagtc atgcccgcga ccccatcttc
781 ggcaaccaga tcatccccga caccgctatc ctcagcgtgg tgccatttca ccacggcttc
841 ggcatgttca ccacgctggg ctacttgatc tgcggctttc gggtcgtgct catgtaccgc
901 ttcgaggagg agctattctt gcgcagcttg caagactata agattcaatc tgccctgctg
961 gtgcccacac tatttagctt cttcgctaag agcactctca tcgacaagta cgacctaagc
1021 aacttgcacg agatcgccag cggcggggcg ccgctcagca aggaggtagg tgaggccgtg
1081 gccaaacgct tccacctacc aggcatccgc cagggctacg gcctgacaga aacaaccagc
1141 gccattctga tcacccccga aggggacgac aagcctggcg cagtaggcaa ggtggtgccc
1201 ttcttcgagg ctaaggtggt ggacttggac accggtaaga cactgggtgt gaaccagcgc
1261 ggcgagctgt gcgtccgtgg ccccatgatc atgagcggct acgttaacaa ccccgaggct
1321 acaaacgctc tcatcgacaa ggacggctgg ctgcacagcg gcgacatcgc ctactgggac
1381 gaggacgagc acttcttcat cgtggaccgg ctgaagagcc tgatcaaata caagggctac
1441 caggtagccc cagccgaact ggagagcatc ctgctgcaac accccaacat cttcgacgcc
1501 ggggtcgccg gcctgcccga cgacgatgcc ggcgagctgc ccgccgcagt cgtcgtgctg
1561 gaacacggta aaaccatgac cgagaaggag atcgtggact atgtggccag ccaggttaca
1621 accgccaaga agctgcgcgg tggtgttgtg ttcgtggacg aggtgcctaa aggactgacc
1681 ggcaagttgg acgcccgcaa gatccgcgag attctcatta aggccaagaa gggcggcaag
1741 atcgccgtga attctgcttg caagaactgg ttcagtagct taagccactt tgtgatccac
1801 cttaacagcc acggcttccc tcccgaggtg gaggagcagg ccgccggcac cctgcccatg
1861 agctgcgccc aggagagcgg catggataga caccctgctg cttgcgccag cgccaggatc
1921 aacgtctaac tgcagtctag agtcggggcg gccggccgct tcgagcagac atgataagat
1981 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
2041 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
2101 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
2161 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tccgtttgcg tattgggcgc
2221 tcttccgctg atctgcgcag caccatggcc tgaaataacc tctgaaagag gaacttggtt
2281 agctaccttc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
2341 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
2401 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
2461 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
2521 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
2581 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
2641 ttgcaaaaag ctcgattctt ctgacactag cgccaccatg accgagtaca agcctaccgt
2701 gcgcctggcc actcgcgatg atgtgccccg cgccgtccgc actctggccg ccgctttcgc
2761 cgactacccc gctacccggc acaccgtgga ccccgaccgg cacatcgagc gtgtgacaga
2821 gttgcaggag ctgttcctga cccgcgtcgg gctggacatc ggcaaggtgt gggtagccga
2881 cgacggcgcg gccgtggccg tgtggactac ccccgagagc gttgaggccg gcgccgtgtt
2941 cgccgagatc ggcccccgaa tggccgagct gagcggcagc cgcctggccg cccagcagca
3001 aatggagggc ctgcttgccc cccatcgtcc caaggagcct gcctggtttc tggccactgt
3061 aggagtgagc cccgaccacc agggcaaggg cttgggcagc gccgtcgtgt tgcccggcgt
3121 agaggccgcc gaacgcgccg gtgtgcccgc ctttctcgaa acaagcgcac caagaaacct
3181 tccattctac gagcgcctgg gcttcaccgt gaccgccgat gtcgaggtgc ccgagggacc
3241 taggacctgg tgtatgacac gaaaacctgg cgcctaatga tctagaaccg gtcatggccg
3301 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg ttcgaactag
3361 atgctgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
3421 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
3481 ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
3541 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
4081 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
4141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
4201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
4261 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
4321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
4381 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagcggcc
4441 gcaaatgcta aaccactgca gtggttacca gtgcttgatc agtgaggcac cgatctcagc
4501 gatctgccta tttcgttcgt ccatagtggc ctgactcccc gtcgtgtaga tcactacgat
4561 tcgtgagggc ttaccatcag gccccagcgc agcaatgatg ccgcgagagc cgcgttcacc
4621 ggcccccgat ttgtcagcaa tgaaccagcc agcagggagg gccgagcgaa gaagtggtcc
4681 tgctactttg tccgcctcca tccagtctat gagctgctgt cgtgatgcta gagtaagaag
4741 ttcgccagtg agtagtttcc gaagagttgt ggccattgct actggcatcg tggtatcacg
4801 ctcgtcgttc ggtatggctt cgttcaactc tggttcccag cggtcaagcc gggtcacatg
4861 atcacccata ttatgaagaa atgcagtcag ctccttaggg cctccgatcg ttgtcagaag
4921 taagttggcc gcggtgttgt cgctcatggt aatggcagca ctacacaatt ctcttaccgt
4981 catgccatcc gtaagatgct tttccgtgac cggcgagtac tcaaccaagt cgttttgtga
5041 gtagtgtata cggcgaccaa gctgctcttg cccggcgtct atacgggaca acaccgcgcc
5101 acatagcagt actttgaaag tgctcatcat cgggaatcgt tcttcggggc ggaaagactc
5161 aaggatcttg ccgctattga gatccagttc gatatagccc actcttgcac ccagttgatc
5221 ttcagcatct tttactttca ccagcgtttc ggggtgtgca aaaacaggca agcaaaatgc
5281 cgcaaagaag ggaatgagtg cgacacgaaa atgttggatg ctcatactcg tcctttttca
5341 atattattga agcatttatc agggttacta gtacgtctct caaggataag taagtaatat
5401 taaggtacgg gaggtattgg acaggccgca ataaaatatc tttattttca ttacatctgt
5461 gtgttggttt tttgtgtgaa tcgatagtac taacatacgc tctccatcaa aacaaaacga
5521 aacaaaacaa actagcaaaa taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc
5581 tct
//