LOCUS       sequence-148262-        6090 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral expression plasmid for GFP with N-terminal 
            BirA-biotinylation signal and TEV celavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6090)
  AUTHORS   Gabriel CH, Gross F, Karl M, Stephanowitz H, Hennig AF, Weber M, 
            Gryzik S, Bachmann I, Hecklau K, Wienands J, Schuchhardt J, Herzel 
            H, Radbruch A, Krause E, Baumgrass R
  TITLE     Identification of Novel Nuclear Factor of Activated T Cells 
            (NFAT)-Associated Proteins in T cells.
  JOURNAL   J Biol Chem. 2016 Sep 16. pii: jbc.M116.739326.
  PUBMED    27637333
REFERENCE   2  (bases 1 to 6090)
  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 Biol 
            Chem. 2016 Sep 16. pii: jbc.M116.739326."
COMMENT     Sequence Label: pMSCV-AVITEV-GFP
FEATURES             Location/Qualifiers
     source          1..6090
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             43..560
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    624..965
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1032..1448
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1378..1400
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1416..1432
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1479..1488
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1497..1541
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1548..1568
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1587..1589,1590..1592,1593..2306)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1632..1653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1893..1912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2240..2261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             2409..2924
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3093..3109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3093..3109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3106..3128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3117..3133
                     /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(3141..3147,3148..3165,3166..3171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3186..3207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3324..3341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3495..4083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3575..3594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4254..5045,5046..5114))
                     /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     4877..4896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5115..5219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5287..5305
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5343..5365)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5465..5484
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5678..5700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5692..5709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5693..5709
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5912..5931
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg
       61 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg
      121 agaatagaga agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca
      181 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
      241 tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
      301 acctgaaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
      361 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggcgcgcc
      421 agtcctccga tagactgcgt cgcccgggta cccgtattcc caataaagcc tcttgctgtt
      481 tgcatccgaa tcgtggactc gctgatcctt gggagggtct cctcagattg attgactgcc
      541 cacctcgggg gtctttcatt tggaggttcc accgagattt ggagacccct gcccagggac
      601 caccgacccc cccgccggga ggtaagctgg ccagcggtcg tttcgtgtct gtctctgtct
      661 ttgtgcgtgt ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact
      721 agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc
      781 ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt
      841 cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac
      901 agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc
      961 tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt
     1021 ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1081 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1141 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1201 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1261 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1321 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1381 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1441 taggcgccgg aattagatct gcggccgcgg atttaaatgc caccatggtt aacagcggcc
     1501 tgaacgatat cttcgaggca cagaaaatcg aatggcatga ggttaacgag aacctgtact
     1561 tccagggcag ctacccggtc gcctccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1621 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg
     1681 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1741 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1801 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1861 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1921 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1981 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2041 atatcatggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     2101 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2161 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2221 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2281 tcggcatgga cgagctgtac aagtaatgaa ttaattaaga attcctgcag cccgggggat
     2341 ccgtcgacct gcagccaagc ttatcgataa aataaaagat tttatttagt ctccagaaaa
     2401 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaatta acgccatttt
     2461 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     2521 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     2581 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     2641 cagatgtttc cagggtgccc caaggacctg aaaatgaccc tgtgccttat ttgaactaac
     2701 caatcagttc gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag
     2761 cccacaaccc ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt
     2821 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     2881 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcatgggtaa cagtttcttg
     2941 aagttggaga acaacattct gagggtagga gtcgaatatt aagtaatcct gactcaatta
     3001 gccactgttt tgaatccaca tactccaata ctcctgaaat agttcattat ggacagcgca
     3061 gaagagctgg ggagaattaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3121 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3181 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3241 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3301 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3901 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     3961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4261 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4381 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4561 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     4621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     4681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     4741 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5161 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5221 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc
     5281 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa
     5341 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     5401 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac
     5461 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     5521 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     5581 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5641 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5701 acggccagtg ccaagctcgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5761 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5821 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5881 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     5941 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgattagt
     6001 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     6061 ccagctgaag cctatagagt acgagccata
//
