LOCUS       sequence-266009-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral control vector containing GFP insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  AUTHORS   Kenific CM, Stehbens SJ, Goldsmith J, Leidal AM, Faure N, Ye J, 
            Wittmann T, Debnath J
  TITLE     NBR1 enables autophagy-dependent focal adhesion turnover.
  JOURNAL   J Cell Biol. 2016 Feb 29;212(5):577-590. Epub 2016 Feb 22.
  PUBMED    26903539
REFERENCE   2  (bases 1 to 6344)
  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: "J Cell 
            Biol."; date: "2016-02-29"; volume: "212(5)"; pages: "577-590. Epub 
            2016 Feb 22"
COMMENT     Sequence Label: pMXspuro-GFP
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..375
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     313..329
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(407..409,410..412,413..1126)
                     /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(407..409,410..412,413..1123)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(452..473)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(713..732)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1060..1081
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1160..1180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1165..1494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1345..1480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1504..2103
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1504..1523)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2000..2020
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             2330..2799
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2929..2946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3100..3688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3180..3199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3852..4643,4644..4712))
                     /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     4475..4494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4713..4817)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             4847..5440
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    5503..5860
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5919..6335
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6265..6287
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6303..6319
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 ccatatgaga tcttatatgg ggcacccccg ccccttgtaa acttccctga ccctgacatg
       61 acaagagtta ctaacagccc ctctctccaa gctcacttac aggctctcta cttagtccag
      121 cacgaagtct ggagacctct ggcggcagcc taccaagaac aactggaccg accggtggta
      181 cctcaccctt accgagtcgg cgacacagtg tgggtccgcc gacaccagac taagaaccta
      241 gaacctcgct ggaaaggacc ttacacagtc ctgctgacca cccccaccgc cctcaaagta
      301 gacggcatcg cagcttggat acacgccgcc cacgtgaagg ctgccgaccc cgggggtgga
      361 ccatcctcta gactgccgga tctagctagt taattaagga tctaccatgg tgagcaaggg
      421 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      481 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      541 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      601 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      661 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      721 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      781 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      841 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      901 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      961 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1021 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1081 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtgactcg agcggccgcc
     1141 agcacagtgg tcgacctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     1201 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     1261 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     1321 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     1381 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     1441 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt
     1501 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     1561 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     1621 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     1681 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     1741 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     1801 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     1861 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     1921 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     1981 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     2041 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     2101 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     2161 tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     2221 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     2281 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg
     2341 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga
     2401 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     2461 ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag
     2521 atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat
     2581 cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca
     2641 caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat
     2701 ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct
     2761 cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt
     2821 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     2881 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3481 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3721 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3781 cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact
     3841 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3901 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3961 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4021 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4081 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4141 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4201 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4261 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4321 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4381 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     4441 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     4501 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     4561 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     4621 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     4681 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     4741 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     4801 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcataatg aaagacccca
     4861 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     4921 aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     4981 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     5041 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     5101 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     5161 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     5221 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     5281 aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     5341 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     5401 ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg tccgggatcg
     5461 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
     5521 tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc gtcggtacta
     5581 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tcggaacacc
     5641 cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg gcccgacctg
     5701 agtccaaaaa tcccgatcgt tttggactct ttggtgcacc cccctaatag gagggatatg
     5761 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
     5821 cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt ctgtgttgtc
     5881 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     5941 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     6001 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     6061 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     6121 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     6181 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     6241 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     6301 ccctttatcc agccctcact ccttctctag gcgcccccat atgg
//
