LOCUS       sequence-101863-        9324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-inactivating retrovirus vector containing a neomycin-resistance
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9324)
  AUTHORS   Fujita T, Fujii H
  TITLE     Identification of Proteins Associated with an IFNgamma-Responsive 
            Promoter by a Retroviral Expression System for enChIP Using CRISPR.
  JOURNAL   PLoS One. 2014 Jul 22;9(7):e103084. doi: 
            10.1371/journal.pone.0103084. eCollection 2014.
  PUBMED    25051498
REFERENCE   2  (bases 1 to 9324)
  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: "PLoS One.";
            date: "2014-07-22"; volume: "9(7):e103084. doi"; pages: " 
            10.1371/journal.pone.0103084. eCollection 2014"
COMMENT     Sequence Label: pSIR-neo
FEATURES             Location/Qualifiers
     source          1..9324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..937
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             996..1412
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1342..1364
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1380..1396
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             4075..4869
                     /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(4129..4148)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4739..4758
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6308..6324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6308..6324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6321..6343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6332..6348
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6356..6362,6363..6380,6381..6386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6401..6422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6539..6556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6710..7298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6790..6809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7469..8260,8261..8329))
                     /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     8092..8111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8330..8434)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8502..8520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8558..8580)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8680..8699
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8907..8924
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8908..8924
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
      421 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
      481 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
      541 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
      601 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
      661 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg
      721 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt
      781 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg
      841 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
      901 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct
      961 gtctgactgt gtttctgtat ttgtctgaga atatgggcca gactgttacc actcccttaa
     1021 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1081 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1141 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1201 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1261 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1321 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1381 tttatccagc cctcactcct tctctaggcg ccaaacctaa acctcaagtt ctttctgaca
     1441 gtggggggcc gctcatcgac ctacttacag aagacccccc gccttatagg gacccaagac
     1501 cacccccttc cgacagggac ggaaatggtg gagaagcgac ccctgcggga gaggcaccgg
     1561 acccctcccc aatggcatct cgcctacgtg ggagacggga gccccctgtg gccgactcca
     1621 ctacctcgca ggcattcccc ctccgcgcag gaggaaacgg acagcttcaa tactggccgt
     1681 tctcctcttc tgacctttac aactggaaaa ataataaccc ttctttttct gaagatccag
     1741 gtaaactgac agctctgatc gagtctgttc tcatcaccca tcagcccacc tgggacgact
     1801 gtcagcagct gttggggact ctgctgaccg gagaagaaaa acaacgggtg ctcttagagg
     1861 ctagaaaggc ggtgcggggc gatgatgggc gccccactca actgcccaat gaagtcgatg
     1921 ccgcttttcc cctcgagttt aaatagtaca taatggattt ccttacgcga aatacgggca
     1981 gacatggcct gcccggttat tattattttt gacaccagac caactggtaa tggtagcgac
     2041 cggcgctcag ctggaattcc gccgatactg acgggctcca ggagtcgtcg ccaccaatcc
     2101 ccatatggaa accgtcgtat aacagccatg tcggttcttc cgcgtgcagc agatggcgat
     2161 ggctggtttc catcagttgc tgttgactgt agcggctgat gttgaactgg aagtcgccgc
     2221 gcccactctg gtgtgggcca taattcaatt cgcgcgtccc gcagcgcaga ccgttttcgc
     2281 tcggaatccc gctcatccgc cacatatctt gatcttcaag ataactcccg tcactccagc
     2341 gcagcaccat caccgcgagg cggttttctc cggcgcgtag atatgcgctg gagctcaaat
     2401 taagacgcca aacgactgtc ttgcccgtaa ccgacccagc gcccgttgca ccacagatga
     2461 aacgccgagt taacgccatc aaaaataatt cgcgtctggc cttcctgtag ccagctttca
     2521 tcaacattaa aatgtgagcg agtaacaacc cgtcggattc tccgtgggaa caaacggcgg
     2581 attgaccgta atgggatagg tcacgttggt gtagatgggc gcatcgtaac cgtgcatctg
     2641 ccagtttgag gggacgacga cagtatcggc ctcaggaaga tcgcactcca gccagctttc
     2701 cggcaccgct tctggtgccg gaaaccaggc aaagcgccat tcgccaattc aggctgcgca
     2761 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     2821 gatgtcctcc aaggccatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     2881 aaacgacggg atcgatctcg ccatacagcg cgttgaaacg ctgggcaata tcgcggctca
     2941 gatcgaggtg ctgtttctgg tcttcaccca ccggtaccag accgccacgg cttacggcaa
     3001 taatgccttt ccattgttca gaaggcatca gtcggcttgc gagtttacgt gcatggatct
     3061 gtcaacatgt cccaggtgac gatgtatttt tcgctcatgt gaagtgtccc agcctgttta
     3121 tctacggctt aaaaagtgtt cgaggggaaa ataggttgcg cgagattata gagatcaagc
     3181 ttgtcgacgg atccgaattc gtcgagggaa gaaataataa agacattaaa atcacttttg
     3241 taaacttttt aaataaggga aattgattaa aacaacgcga agttgtaaaa tgccatatac
     3301 tgcatacatt ttgtgggaaa aaatgtgcat ttacctgctc accaaattat cttattagct
     3361 agcagtagag ctctttgtgt gatgggaaga tgggataaca gctgttgtta ctgtactggt
     3421 tttttttttt aatatattta aaaaaaagag tgtctcctaa agtgtaagcg ctatcatgct
     3481 ggactaatgt tttaatttaa tttaattgtt gtcttggtag agatggggtc tcgctatttt
     3541 gctcacgcag atctcaaact cctgacgccc aggtacactc ccatctcggc ctcctaaaat
     3601 ggtgggatca cagacgtcag ccaccgagcc cggtcacttt tttgtattcc ccacagtatt
     3661 gatgtatatc ttctgcgttc aaaagcaatt ttttaaagcc tcataacgtg gtaacagaat
     3721 actttgcaca ttacaaaatt cagaacacgg aaacaagaag ctcgcttttt tttcccccct
     3781 atttcggttt ggccctttag atttcccctc ccccaccggg gcgggacttc ccgccgactt
     3841 ctttcaggtt ctcagttcgg tccgccaact gtcgtataaa ggcgctgcct caggccagag
     3901 gcctcacaaa gcgcgtcgag acagcaagcg aaccggaatt gccagctggg gcgccctctg
     3961 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
     4021 ggcgcagggg atcaagatcg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4081 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4141 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4201 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4261 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4321 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4381 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4441 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4501 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4561 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4621 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4681 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4741 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4801 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4861 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4921 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     4981 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagatc ccgcgattag
     5041 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     5101 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     5161 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     5221 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     5281 gaacagatgg aacaggtcga cctagagaac catcagatgt ttccagggtg ccccaaggac
     5341 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     5401 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     5461 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     5521 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     5581 agcgggggtc tcttcacatg cagcatgtat caaaattaat ttggtttttt tttcttaagt
     5641 atttacatta aatggcccat agtacttaaa gtaacatggg cttccttgaa ataaacatgg
     5701 agtattcaga atgtgtcata aatatttcta attttaagat agtatctcca ttggctatct
     5761 actttttctt atattttttt ttgtcctctg tcttccattc gttgttgttg ttgtttgttt
     5821 gtttgtttgt tggttggttg gttaattttt ttttaaagat cctacactat agttcaagct
     5881 agactattag ctactctgta acccagggtg acctttgaag tcatgggtag cctgctgttt
     5941 tagccttccc acatctaaga ttacaggtat gagctatcat ttttggtata ttgattgatt
     6001 gattgattga tgtgtgtgtg tgtgattgtg tttgtgtgtg tgattgtgta tatgtgtgta
     6061 tggttgtgtg tgattgtgtg tatgtatgtt tgtgtgtgat tgtgtgtgtg tgattgtgca
     6121 tgtgtgtgtg tgtgattgtg tttatgtgta tgattgtgtg tgtgtgtgtg tgtgtgtgtg
     6181 tgtgtgtgtg tgtgtgtgtg ttgtgtatat atatttatgg tagtgagagg caacgctccg
     6241 gctcaggtgt caggttggtt tttgagacag agtctttcgc ttagcttgga attaattcgt
     6301 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6361 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6421 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6481 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6541 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6721 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6781 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6841 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6901 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6961 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7021 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7081 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7141 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7201 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7261 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7321 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7381 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7441 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7501 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7561 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7621 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7681 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7741 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7801 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7861 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7921 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7981 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     8041 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8101 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8161 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8221 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8281 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     8341 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8401 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8461 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     8521 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     8581 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     8641 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     8701 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     8761 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8821 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     8881 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag ctagctttgc
     8941 tcttaggagt ttcctaatac atcccaaact caaatatata aagcatttga cttgttctat
     9001 gccctagggg gcggggggaa gctaagccag ctttttttaa catttaaaat gttaattcca
     9061 ttttaaatgc acagatgttt ttatttcata agggtttcaa tgtgcatgaa tgctgcaata
     9121 ttcctgttac caaagctagt ataaataaaa atagataaac gtggaaatta cttagagttt
     9181 ctgtcattaa cgtttccttc ctcagttgac aacataaatg cgctgctgag aagccagttt
     9241 gcatctgtca ggatcaattt cccattatgc cagtcatatt aattactagt caattagttg
     9301 atttttattt ttgacatata catg
//
