LOCUS       sequence-356572-        7005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of Cdc42 splice variant 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7005)
  AUTHORS   Rehn M, Wenzel A, Frank AK, Schuster MB, Pundhir S, Jorgensen N, 
            Vitting-Seerup K, Ge Y, Jendholm J, Michaut M, Schoof EM, Jensen TL,
            Rapin N, Sapio RT, Andersen KL, Lund AH, Solimena M, Holzenberger M,
            Pestov DG, Porse BT
  TITLE     PTBP1 promotes hematopoietic stem cell maintenance and red blood 
            cell development by ensuring sufficient availability of ribosomal 
            constituents.
  JOURNAL   Cell Rep. 2022 May 10;39(6):110793. doi: 
            10.1016/j.celrep.2022.110793.
  PUBMED    35545054
REFERENCE   2  (bases 1 to 7005)
  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: "Cell Rep.";
            date: "2022-05-10"; volume: "39(6):110793. doi"; pages: " 
            10.1016/j.celrep.2022.110793"
COMMENT     Sequence Label: MIGR1-Cdc42-2
FEATURES             Location/Qualifiers
     source          1..7005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     147..166
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     406..427
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    484..1036
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    486..1053
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(646..665)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     875..892
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(1646..2062)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(1662..1678)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1694..1716)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2129..2470
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             2534..3050
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3250..3269)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(3397..3419)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3613..3632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3732..3754
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3792..3810)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3878..3982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3983..4051,4052..4843)
                     /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     complement(4201..4220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5014..5602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5503..5522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5756..5773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5890..5911
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5926..5931,5932..5949,5950..5956)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5964..5980
                     /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)."
     LTR             6142..6656
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(join(6758..7005,1..466,467..469,470..472))
                     /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(6803..6824)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
       61 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
      121 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
      181 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
      241 ggtcggggta gcggctgaag cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc
      301 agggcacggg cagcttgccg gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg
      361 catcgccctc gccctcgcca gacacgctga acttgtggcc gtttacgtcg ccgtccagct
      421 cgaccaggat gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggttgtgg
      481 ccatattatc atcgtgtttt tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga
      541 cgttttttta acctcgacta aacacatgta aagcatgtgc accgaggccc cagatcagat
      601 cccatacaat ggggtacctt ctgggcatcc ttcagcccct tgttgaatac gcttgaggag
      661 agccatttga ctctttccac aactatccaa ctcacaacgt ggcactgggg ttgtgccgcc
      721 tttgcaggtg tatcttatac acgtggcttt tggccgcaga ggcacctgtc gccaggtggg
      781 gggttccgct gcctgcaaag ggtcgctaca gacgttgttt gtcttcaaga agcttccaga
      841 ggaactgctt ccttcacgac attcaacaga ccttgcattc ctttggcgag aggggaaaga
      901 cccctaggaa tgctcgtcaa gaagacaggg ccaggtttcc gggccctcac attgccaaaa
      961 gacggcaata tggtggaaaa taacatatag acaaacgcac accggcctta ttccaagcgg
     1021 cttcggccag taacgttagg gggggggggc ggaattctta gaatatacag cacttccttt
     1081 tgggttgagt ttccggaggc tcgagggcag ctaggatagc ctcatcaaac acattcttca
     1141 gacctctctg tgtaagtgca gaacactcca catacttgac agccttcagg tcacgggcca
     1201 gcttttcagc agtctctgga gtgataggct tctgtttgtt cttggcaagt ttctcaatag
     1261 tagaggggtc atctctgaga tcaatttgag tcccaacaag caagaaagga gtctttggac
     1321 agtggtgagt tatctcaggc acccactttt ctttcacgtt ttcaaatgaa gatggagaga
     1381 ccactgaaaa acagactaga aatacatctg tttgtggata actcagcggt cgtaatctgt
     1441 cataatcctc ttgccctgca gtatcaaaaa gtccaagagt atatggttct ccaccaatca
     1501 taactgtgac tgcatagttg tcaaaaacag tcggtacata ttccgatgga aatttgtttg
     1561 ttgtgtagga tatcaggaga catgttttac caacagcacc atcgcccaca acaacacact
     1621 taattgtctg catagatcta attccggcgc ctagagaagg agtgagggct ggataaaggg
     1681 aggatcgagg cggggtcgaa cgaggaggtt caagggggag agacggggcg gatggaggaa
     1741 gaggaggcgg aggcttaggg tgtacaaagg gcttgaccca gggagggggg tcaaaagcca
     1801 aggcttccca ggtcacgatg taggggacct ggtctgggtg tccatgcggg ccaggtgaaa
     1861 agaccttgat cttaacctgg gtgatgaggt ctcggttaaa ggtgccgtct cgcggccatc
     1921 cgacgttaaa ggttggccat tctgcagagc agaaggtaac ccaacgtctc ttcttgacat
     1981 ctaccgactg gttgtgagcg atccgctcga catctttcca gtgacctaag gtcaaactta
     2041 agggagtggt aacagtctgg ccctaatttt cagacaaata cagaaacaca gtcagacaga
     2101 gacaacacag aacgatgctg cagcgctgca gcagacaaga cgcgcggctt cggttccaaa
     2161 ccgaaagcaa aaattcagac ggaggcggga actgttttag gttctcgtct cctaccagaa
     2221 ccacatatcc tgacggggtc ggattccaca tcgactccct tcctcaggtc gggccacaaa
     2281 aacggccccc aaagtccctg ggacgtctcc cagggttgcg gccgggtgtt cagaactcgt
     2341 cagttccacc acgggtccgc cagatacaga gctagttagc taactagtac agacgcaggc
     2401 gcaaacatta gatgccggca caaacacgca caaagacaga gacagacacg aaacgaccgc
     2461 tggccagctt acctcccggc gggggggtcg gtggtccctg ggcaggggtc tccaaatctc
     2521 ggtggaacct ccaaatgaaa gacccccgag gtgggcagtc aatcaatctg aggagaccct
     2581 cccaaggatc agcgagtcca cgattcggat gcaaacagca agaggcttta ttgggaatac
     2641 gggtacccgg gcgacgcagt ctatcggagg actggcgcgc cgagtgaggg gttgtgggct
     2701 cttttattga gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt
     2761 tagttcaaat aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg
     2821 gttctctaga aactgctgag ggcgggaccg catctgggga ccatctgttc ttggccctga
     2881 gccggggcag gaactgctta ccacagatat cctgtttggc ccatattctg ctgtctctct
     2941 gttcctaacc ttgatctgaa cttctctatt ctcagttatg tattttccat gccttgcaaa
     3001 atggcgttac ttaagctagc ttgccaaacc tacaggtggg gtctttcatt cccccctttt
     3061 tctggagact aaataaaatc ttttatttta tctatggctc gtactctata ggcttcagct
     3121 ggtgatattg ttgagtcaaa actagagcct ggaccactga tatcctgtct ttaacaaatt
     3181 ggactaatcg cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     3241 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     3301 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     3361 gcgccatctc cttgcatgca ccattccttg cgccgtcgtt ttacaacgtc gtgactggga
     3421 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3481 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3541 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     3601 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3661 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3721 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3781 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3841 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3901 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3961 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4021 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4081 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4141 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4201 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4261 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4321 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4381 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4441 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4501 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4561 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     4621 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4681 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4741 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4801 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4861 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4921 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4981 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5041 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5101 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5161 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5221 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5281 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5341 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5401 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5461 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5521 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5581 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5641 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5701 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5761 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     5821 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5881 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     5941 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacaatt ctccccagct
     6001 ctttctgcgc tgtccataat gaactatttc aggagtattg gagtatgtgg attcaaaaca
     6061 gtggctaatt gagtcaggat tacttaatat tcgactccta ccctcagaat gttgttctcc
     6121 aacttcaaga aactgttacc catgaaagac ccccgctgac gggtagtcaa tcactcagag
     6181 gagaccctcc caaggaacag cgagaccaca agtcggatgc aactgcaaga gggtttattg
     6241 gatacacggg tacccgggcg acgcagtcta tcggaggact ggcgcgccga gtgaggggtt
     6301 gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga gaagcgaact
     6361 gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac cctggaaaca
     6421 tctgatggtt ctctagaaac tgctgagggc gggaccgcat ctggggacca tctgttcttg
     6481 gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca tattctgctg
     6541 tctctctgtt cctaaccttg atctgaactt ctctattctc agttatgtat tttccatgcc
     6601 ttgcaaaatg gcgttactta agctagcttg ccaaacctac aggtggggtc tttcattccc
     6661 ccctttttct ggagactaaa taaaatcttt tattttatcg ataagcttgg ctgcaggtcg
     6721 acggtatcga taagcttgat aattcttaat taattcatta cttgtacagc tcgtccatgc
     6781 cgagagtgat cccggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     6841 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     6901 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ctgccgtcct
     6961 cgatgttgtg gcggatcttg aagttcgcct tgatgccgtt cttct
//
