LOCUS       sequence-11577-e        6582 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 6582)
  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 6582)
  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..6582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1816..1839
                     /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 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc ctcgagaatt catgatggat caagccagat cagcattctc taacttgttt
     1141 ggtggggaac cattgtcata cacccggttt agccttgctc ggcaagtaga tggagataac
     1201 agtcatgtgg agatgaaact ggctgcagat gaagaagaaa atgccgacaa taacatgaag
     1261 gctagtgtca gaaaacccaa gaggtttaat ggaagactct gctttgcagc tattgcacta
     1321 gtcattttct tcttgattgg attcatgagt ggctacctgg gctattgtaa gcgtgtagaa
     1381 caaaaagagg agtgtgtgaa actggctgaa acggaggaga cagacaagtc agaaaccatg
     1441 gaaacagagg atgttgctgc agatcaagcc agagagctca tcaattcctg ggtagaaagt
     1501 cagacaaatg gaattatcag aaatgtcctt cagccaagct ccgtggattc tcaaactgca
     1561 atggttctgg ttaatgccat tgtcttcaaa ggactgtggg agaaaacatt taaggatgaa
     1621 gacacacaag caatgccttt cagagtgact gagcaagaaa gcaaacctgt gcagatgatg
     1681 taccagattg gtttatttag agtggcatca atggcttctg agaaaatgaa gatcctggag
     1741 cttccatttg ccagtgggac aatgagcatg ttggtgctgt tgcctgatga agtctcaggc
     1801 cttgagcagc ttgagagtat aatcaacttt gaaaaactga ctgaatggac cagttctaat
     1861 gttatggaag agaggaagat caaagtgtac ttacctcgca tgaagatgga ggaaaaatac
     1921 aacctcacat ctgtcttaat ggctatgggc attactgacg tgtttagctc ttcagccaat
     1981 ctgtctggca tctcctcagc agagagcctg aagatatctc aagctgtcca tgcagcacat
     2041 gcagaaatca atgaagcagg cagagaggtg gtagggtcag cagaggctgg agtggatgct
     2101 gcaagcgtct ctgaagaatt tagggctgac catccattcc tcttctgtat caagcacatc
     2161 gcaaccaacg ccgttctctt ctttggcaga tgtgtttccc cttaagaatt 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 ct
//
