LOCUS       sequence-464634-        6950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Control retroviral vector expressing Green Fluorescent Protein. 
            Neomycin selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6950)
  AUTHORS   Addgene Research Program
  TITLE     Addgene Retroviral Backbones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6950)
  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            "Unpublished"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 EGFP Neo
FEATURES             Location/Qualifiers
     source          1..6950
                     /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"
     CDS             join(1453..1455,1456..1458,1459..2172)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1453..1455,1456..1458,1459..2169)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1498..1519)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1759..1778)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2106..2127
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2209..2229)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2214..2543
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2394..2529
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2553..3347
                     /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(2607..2626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3217..3236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             3568..4037
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4143..4163)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4143..4161)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4182..4198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4182..4198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4195..4217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4206..4222
                     /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(4230..4236,4237..4254,4255..4260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4275..4296
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4413..4430)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4584..5172)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4664..4683)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5343..6134,6135..6203))
                     /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     5966..5985
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6204..6308)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6334..6789)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6471..6492)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6683..6702
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6916..6938
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6930..6947
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6931..6947
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1501 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1561 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1621 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     1681 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1741 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1801 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1861 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1921 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1981 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     2041 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     2101 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2161 ctgtacaagt aatacgtagg aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc
     2221 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2281 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2341 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2401 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2461 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2521 ttggaggcct aggcttttgc aaaaagctta ccatgattga acaagatgga ttgcacgcag
     2581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2701 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3061 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgacgc ccgccccacg
     3361 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa taaaagattt
     3421 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     3481 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     3541 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     3601 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     3661 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     3721 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     3781 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     3841 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     3901 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     3961 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4021 tcagcggggg tctttcacat gcagcatgta tcggcgccaa gccgaattct gcagatatcc
     4081 atcacactgg cggccgctcg actagagcgg ccgccaccgc ggtggagctc cagcttttgt
     4141 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     4201 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4261 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4321 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4381 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4441 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4501 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4561 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4621 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4681 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4741 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4801 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4861 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4921 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4981 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5041 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5101 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5161 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5221 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5281 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5341 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5401 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5461 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5521 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5581 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5641 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5701 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5761 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5821 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5881 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5941 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6001 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6061 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6121 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6181 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6241 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6301 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     6361 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6421 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6481 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6541 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6601 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6661 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6721 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6781 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     6841 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     6901 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
//
