LOCUS       sequence-464633-        6241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for cloning and expressing your gene of interest. 
            Neomycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6241)
  AUTHORS   Addgene Research Program
  TITLE     Addgene Retroviral Backbones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6241)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAG Retro Neo Empty
FEATURES             Location/Qualifiers
     source          1..6241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     50..66
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     51..67
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             92..561
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    626..825
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1026..1442
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1372..1394
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1410..1426
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1500..1520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1505..1834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1685..1820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1844..2638
                     /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(1898..1917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2508..2527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             2859..3328
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3434..3454)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3434..3452)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3473..3489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3473..3489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3486..3508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3497..3513
                     /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(3521..3527,3528..3545,3546..3551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3566..3587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3704..3721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3875..4463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3955..3974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4634..5425,5426..5494))
                     /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     5257..5276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5495..5599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5625..6080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5762..5783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5974..5993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6207..6229
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6221..6238
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6222..6238
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
       61 ggtatcgggg gcatgcagtg ccacctgcag cctgaatatg ggccaaacag gatatctgtg
      121 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
      181 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
      241 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
      301 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
      361 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
      421 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
      481 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
      541 cccgtcagcg ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg
      601 gaccaccgac ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt ctgtccgatt
      661 gtctagtgtc tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct
      721 gtatctggcg gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga
      781 gacgtcccag ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt
      841 ggaatccgac cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
      901 cgcctccgtc tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc
      961 agcgctgcag catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa
     1021 attagggcca gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg
     1081 agcggatcgc tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct
     1141 ctgcagaatg gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc
     1201 tcatcaccca ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg
     1261 tcccctacat cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct
     1321 ttgtacaccc taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac
     1381 ctcctcgttc gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg
     1441 ccggccggat cccagtgtgg tggtacgtag gaattcgcca gcacagtggt cgaccctgtg
     1501 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1561 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     1621 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     1681 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1741 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     1801 aggaggcttt tttggaggcc taggcttttg caaaaagctt accatgattg aacaagatgg
     1861 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     1921 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     1981 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2041 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2101 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     2161 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     2221 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     2281 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     2341 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     2401 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     2461 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     2521 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     2581 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgacg
     2641 cccgccccac gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgataaa
     2701 ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt
     2761 tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat
     2821 agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat
     2881 atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata
     2941 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     3001 tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg
     3061 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     3121 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     3181 tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc
     3241 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     3301 ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcggcgcca agccgaattc
     3361 tgcagatatc catcacactg gcggccgctc gactagagcg gccgccaccg cggtggagct
     3421 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     3481 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3541 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3601 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3661 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3721 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3781 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3841 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3901 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3961 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4021 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4081 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4141 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4201 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4261 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4321 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4381 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4441 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4501 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4561 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4621 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4681 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4741 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4801 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4861 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4921 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4981 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5041 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5101 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5161 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5221 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5281 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5341 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5401 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5461 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5521 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5581 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5641 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5701 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5761 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5821 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5881 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5941 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     6001 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     6061 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     6121 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     6181 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     6241 g
//
