LOCUS pCI-neo-mOVA 6594 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCI-neo-mOVA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6594)
AUTHORS Yang J, Sanderson NS, Wawrowsky K, Puntel M, Castro MG, Lowenstein
PR
TITLE Kupfer-type immunological synapse characteristics do not predict
anti-brain tumor cytolytic T-cell function in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2010 Mar 9. 107(10):4716-21.
PUBMED 20133734
REFERENCE 2 (bases 1 to 6594)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6594
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 126..505
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 506..717
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 668..688
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 714..738
/label=LNCX
/note="Human CMV promoter, forward primer"
intron 878..1010
/label=chimeric intron
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
primer_bind 993..1012
/label=pMT2-F
/note="Synthetic intron, forward primer"
promoter 1055..1073
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1810..1833
/codon_start=1
/product="antigenic CD8+ T cell epitope from chicken
ovalbumin"
/label=OVA peptide
/translation="SIINFEKL"
promoter complement(2251..2268)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 2286..2407
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2297..2316)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2351..2370
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
rep_origin 2593..3048
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2680..2699)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2890..2911
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3165..3522
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(3188..3208)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3373..3508
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3435..3454
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3573..4367
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3627..3646)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4237..4256
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4431..4479
/note="synthetic polyadenylation signal"
primer_bind complement(4520..4539)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4639..4661
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(4699..4717)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 4785..4889
/gene="bla"
/label=AmpR promoter
CDS 4890..5750
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5108..5127)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5921..6509
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6410..6429
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
61 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
121 atgttggcat tgattattga ctagttatta atagtaatca attacggggt cattagttca
181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
361 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
421 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
601 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactgcg atcgcccgcc
661 ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
721 ttagtgaacc gtcagatcac tagaagcttt attgcggtag tttatcacag ttaaattgct
781 aacgcagtca gtgcttctga cacaacagtc tcgaacttaa gctgcagtga ctctcttaag
841 gtagccttgc agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca
901 ggtttaagga gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg
961 ataggcacct attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc
1021 ccagttcaat tacagctctt aaggctagag tacttaatac gactcactat aggctagcct
1081 cgagaattcg tcgacatgat ggatcaagcc agatcagcat tctctaactt gtttggtggg
1141 gaaccattgt catacacccg gtttagcctt gctcggcaag tagatggaga taacagtcat
1201 gtggagatga aactggctgc agatgaagaa gaaaatgccg acaataacat gaaggctagt
1261 gtcagaaaac ccaagaggtt taatggaaga ctctgctttg cagctattgc actagtcatt
1321 ttcttcttga ttggattcat gagtggctac ctgggctatt gtaagcgtgt agaacaaaaa
1381 gaggagtgtg tgaaactggc tgaaacggag gagacagaca agtcagaaac catggaaaca
1441 gaggatgttg ctgcagatca agccagagag ctcatcaatt cctgggtaga aagtcagaca
1501 aatggaatta tcagaaatgt ccttcagcca agctccgtgg attctcaaac tgcaatggtt
1561 ctggttaatg ccattgtctt caaaggactg tgggagaaaa catttaagga tgaagacaca
1621 caagcaatgc ctttcagagt gactgagcaa gaaagcaaac ctgtgcagat gatgtaccag
1681 attggtttat ttagagtggc atcaatggct tctgagaaaa tgaagatcct ggagcttcca
1741 tttgccagtg ggacaatgag catgttggtg ctgttgcctg atgaagtctc aggccttgag
1801 cagcttgaga gtataatcaa ctttgaaaaa ctgactgaat ggaccagttc taatgttatg
1861 gaagagagga agatcaaagt gtacttacct cgcatgaaga tggaggaaaa atacaacctc
1921 acatctgtct taatggctat gggcattact gacgtgttta gctcttcagc caatctgtct
1981 ggcatctcct cagcagagag cctgaagata tctcaagctg tccatgcagc acatgcagaa
2041 atcaatgaag caggcagaga ggtggtaggg tcagcagagg ctggagtgga tgctgcaagc
2101 gtctctgaag aatttagggc tgaccatcca ttcctcttct gtatcaagca catcgcaacc
2161 aacgccgttc tcttctttgg cagatgtgtt tccccttaag tcgacgaatt cacgcgtggt
2221 acctctagag tcgacccggg cggccgcttc cctttagtga gggttaatgc ttcgagcaga
2281 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
2341 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
2401 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
2461 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atccgataag gatcgatccg
2521 ggctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
2581 aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
2641 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
2701 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
2761 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
2821 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
2881 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
2941 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
3001 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
3061 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
3121 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
3181 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
3241 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
3361 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
3421 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
3481 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac
3541 acaacagtct cgaacttaag gctagagcca ccatgattga acaagatgga ttgcacgcag
3601 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
3661 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
3721 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
3781 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
3841 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
3901 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
3961 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
4021 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
4081 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
4141 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
4201 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
4261 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
4321 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
4381 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat
4441 ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc
4501 gcgtatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
4561 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
4621 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
4681 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
4741 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
4801 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
4861 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
4921 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
4981 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
5041 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
5101 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
5161 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
5221 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
5281 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
5341 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
5401 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
5461 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
5521 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
5581 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
5641 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
5701 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
5761 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
5821 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
5881 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
5941 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
6001 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
6061 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
6121 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
6181 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
6241 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
6301 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
6361 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
6421 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
6481 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
6541 tggccttttg ctggcctttt gctcacatgg ctcgacagat cttcaatatt ggcc
//
