LOCUS       sequence-197341-        6594 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 2010 Mar 9. 107(10):4716-21."
COMMENT     Sequence Label: pCI-neo-mOVA
FEATURES             Location/Qualifiers
     source          1..6594
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     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"
     enhancer        3165..3356
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     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"
     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             join(4890..4958,4959..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
//
