LOCUS       sequence-219895-        6765 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 6765)
  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 6765)
  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-sOVA
FEATURES             Location/Qualifiers
     source          1..6765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        144..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..735
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     686..706
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     732..756
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          896..1028
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1011..1030
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1073..1091
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1133..2293
                     /codon_start=1
                     /product="chicken ovalbumin"
                     /label=ovalbumin
                     /translation="MGSIGAASMEFCFDVFKELKVHHANENIFYCPIAIMSALAMVYLG
                     AKDSTRTQINKVVRFDKLPGFGDSIEAQCGTSVNVHSSLRDILNQITKPNDVYSFSLAS
                     RLYAEERYPILPEYLQCVKELYRGGLEPINFQTAADQARELINSWVESQTNGIIRNVLQ
                     PSSVDSQTAMVLVNAIVFKGLWEKAFKDEDTQAMPFRVTEQESKPVQMMYQIGLFRVAS
                     MASEKMKILELPFASGTMSMLVLLPDEVSGLEQLESIINFEKLTEWTSSNVMEERKIKV
                     YLPRMKMEEKYNLTSVLMAMGITDVFSSSANLSGISSAESLKISQAVHAAHAEINEAGR
                     EVVGSAEAGVDAASVSEEFRADHPFLFCIKHIATNAVLFFGRCVSP"
     polyA_signal    2475..2596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2486..2505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2540..2559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2782..3237
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2869..2888)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3079..3100
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3354..3711
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3354..3545
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3377..3397)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3562..3697
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3762..4556
                     /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(3816..3835)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4426..4445
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4620..4668
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4709..4728)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4828..4850
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4888..4906)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4974..5078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5079..5147,5148..5939)
                     /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(5297..5316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6110..6698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6599..6618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatcttcaa tattggccat tagccatatt attcattggt tatatagcat aaatcaatat
       61 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
      121 atgtccaata tgaccgccat gttggcattg attattgact agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactgcgat cgcccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      721 tatataagca gagctcgttt agtgaaccgt cagatcacta gaagctttat tgcggtagtt
      781 tatcacagtt aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc
      841 tgcagtgact ctcttaaggt agccttgcag aagttggtcg tgaggcactg ggcaggtaag
      901 tatcaaggtt acaagacagg tttaaggaga ccaatagaaa ctgggcttgt cgagacagag
      961 aagactcttg cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct
     1021 ctccacaggt gtccactccc agttcaatta cagctcttaa ggctagagta cttaatacga
     1081 ctcactatag gctagcctcg agaattcctc gaaagacaac tcagagttca ccatgggctc
     1141 catcggtgca gcaagcatgg aattttgttt tgatgtattc aaggagctca aagtccacca
     1201 tgccaatgag aacatcttct actgccccat tgccatcatg tcagctctag ccatggtata
     1261 cctgggtgca aaagacagca ccaggacaca aataaataag gttgttcgct ttgataaact
     1321 tccaggattc ggagacagta ttgaagctca gtgtggcaca tctgtaaacg ttcactcttc
     1381 acttagagac atcctcaacc aaatcaccaa accaaatgat gtttattcgt tcagccttgc
     1441 cagtagactt tatgctgaag agagataccc aatcctgcca gaatacttgc agtgtgtgaa
     1501 ggaactgtat agaggaggct tggaacctat caactttcaa acagctgcag atcaagccag
     1561 agagctcatc aattcctggg tagaaagtca gacaaatgga attatcagaa atgtccttca
     1621 gccaagctcc gtggattctc aaactgcaat ggttctggtt aatgccattg tcttcaaagg
     1681 actgtgggag aaagcattta aggatgaaga cacacaagca atgcctttca gagtgactga
     1741 gcaagaaagc aaacctgtgc agatgatgta ccagattggt ttatttagag tggcatcaat
     1801 ggcttctgag aaaatgaaga tcctggagct tccatttgcc agtgggacaa tgagcatgtt
     1861 ggtgctgttg cctgatgaag tctcaggcct tgagcagctt gagagtataa tcaactttga
     1921 aaaactgact gaatggacca gttctaatgt tatggaagag aggaagatca aagtgtactt
     1981 acctcgcatg aagatggagg aaaaatacaa cctcacatct gtcttaatgg ctatgggcat
     2041 tactgacgtg tttagctctt cagccaatct gtctggcatc tcctcagcag agagcctgaa
     2101 gatatctcaa gctgtccatg cagcacatgc agaaatcaat gaagcaggca gagaggtggt
     2161 agggtcagca gaggctggag tggatgctgc aagcgtctct gaagaattta gggctgacca
     2221 tccattcctc ttctgtatca agcacatcgc aaccaacgcc gttctcttct ttggcagatg
     2281 tgtttcccct taaaaagaag aaagctgaaa aactctgtcc cttccaacaa gacccagagc
     2341 actgtagtat caggggtaaa atgaaaagta tgttatctgc tgcatccaga cttcataaaa
     2401 gctggagctt aatctagagt cgacccgggc ggccgcttcc ctttagtgag ggttaatgct
     2461 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2521 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2581 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2641 gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tccgataagg
     2701 atcgatccgg gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2761 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     2821 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     2881 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2941 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3001 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3061 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3121 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3181 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct
     3241 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gcggatctgc
     3301 gcagcaccat ggcctgaaat aacctctgaa agaggaactt ggttaggtac cttctgaggc
     3361 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3421 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3481 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3541 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3601 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3661 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgat
     3721 tcttctgaca caacagtctc gaacttaagg ctagagccac catgattgaa caagatggat
     3781 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3841 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3901 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3961 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4021 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4081 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4141 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4201 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4261 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     4321 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4381 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4441 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4501 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4561 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     4621 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     4681 taaggatccg cgtatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4741 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4801 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4861 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4921 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4981 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5041 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5101 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5161 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5221 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5281 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5341 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5401 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5461 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5521 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5581 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5641 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5701 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5761 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5821 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5881 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5941 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6001 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6061 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6121 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6181 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6241 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6301 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6361 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6421 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6481 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6541 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6601 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6661 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6721 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgac
//
