LOCUS Exported 4531 bp ds-DNA circular SYN 19-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..4531
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 67..370
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 371..574
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 620..638
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 682..759
/codon_start=1
/product="calmodulin-binding peptide"
/label=CBP
/note="derived from skeletal muscle myosin light chain
kinase; binds calmodulin with nanomolar affinity in the
presence of calcium"
/translation="KRRWKKNFIAVSAANRFKKISSSGAL"
CDS 778..891
/codon_start=1
/product="streptavidin-binding peptide"
/label=SBP
/note="selected from a peptide library; binds streptavidin
with nanomolar affinity (Keefe et al., 2001)"
/translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
misc_feature 914..987
/label=MCS
promoter complement(1033..1051)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1325..1446
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(1453..1908)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 1935..2039
/gene="bla"
/label=AmpR promoter
promoter 2041..2398
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2249..2384
/label=SV40 ori
/note="SV40 origin of replication"
CDS 2433..3227
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3459..3506
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 3835..4423
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
661 gcggtggcgg ccgccaccat gaagcgacga tggaaaaaga atttcatagc cgtctcagca
721 gccaaccgct ttaagaaaat ctcatcctcc ggggcacttg gaagcggtag cggtaccatg
781 gacgagaaga ccaccggctg gcggggcggc cacgtggtgg agggcctggc cggcgagctg
841 gagcagctgc gggccaggct ggagcaccac cctcagggcc agcgggagcc ctccggcggc
901 tgcaagctgg gctgcccggg cggatccccc gggctgcagg aattcgatat caagcttatc
961 gataccgtcg acctcgaggg ggggcccggt accttaatta attaaggtac caggtaagtg
1021 tacccaattc gccctatagt gagtcgtatt acaattcact cgatcgccct tcccaacagt
1081 tgcgcagcct gaatggcgaa tggagatcca atttttaagt gtataatgtg ttaaactact
1141 gattctaatt gtttgtgtat tttagattca cagtcccaag gctcatttca ggcccctcag
1201 tcctcacagt ctgttcatga tcataatcag ccataccaca tttgtagagg ttttacttgc
1261 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
1321 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
1381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
1441 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
1501 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
1561 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
1621 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
1681 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
1741 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
1801 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
1861 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
1921 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
1981 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagaatc
2041 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
2101 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
2161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
2221 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
2281 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
2341 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
2401 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
2461 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
2521 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
2581 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
2641 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
2701 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
2761 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
2821 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
2881 ccggtcttgt cgatcaggat gatctggacg aagaacatca ggggctcgcg ccagccgaac
2941 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
3001 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
3061 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
3121 aagaacttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
3181 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
3241 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
3301 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
3361 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
3421 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
3481 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
3541 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
3601 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
3661 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
3721 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
3781 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
3841 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
3901 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
3961 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
4021 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
4081 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
4141 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
4201 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
4261 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
4321 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
4381 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
4441 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
4501 ccctgattct gtggataacc gtattaccgc c
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