LOCUS       sequence-439160-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression vector for an shDNM1L.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  AUTHORS   Sweatt AJ, Griffiths CD, Groves SM, Paudel BB, Wang L, Kashatus DF, 
            Janes KA
  TITLE     Proteome-wide copy-number estimation from transcriptomics.
  JOURNAL   Mol Syst Biol. 2024 Sep 27. doi: 10.1038/s44320-024-00064-3.
  PUBMED    39333715
REFERENCE   2  (bases 1 to 7453)
  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: "Mol Syst 
            Biol. 2024 Sep 27. doi: 10.1038/s44320-024-00064-3."
COMMENT     Sequence Label: pSuperior.retro.neo GFP shDNM1L
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(541..560)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(688..710)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(904..923)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1023..1045
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1083..1101)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1169..1273
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1274..1342,1343..2134)
                     /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(1492..1511)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2305..2893
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2794..2813
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3047..3064
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3181..3202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3217..3222,3223..3240,3241..3247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3255..3271
                     /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)."
     polyA_signal    3832..3880
                     /note="synthetic polyadenylation signal"
     CDS             complement(3944..4738)
                     /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(4055..4074)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4665..4684
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(join(4802..5512,5513..5515,5516..5518))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(4802..5512,5513..5515,5516..5518))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4844..4865)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5193..5212
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5452..5473
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5515..5524
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5553..6056)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(5601..5620)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     5998..6021
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     6202..6221
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    6267..6285
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(6355..6371)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6356..6372)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(6390..6806)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6406..6422)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6438..6460)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    6873..7214
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             join(7278..7453,1..341)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 agctcgggga gcagaagcgc gcgaacagaa gcgagaagcg aactgattgg ttagttcaaa
       61 taaggcacag ggtcatttca ggtccttggg gcaccctgga aacatctgat ggttctctag
      121 aaactgctga gggcgggacc gcatctgggg accatctgtt cttggccctg agccggggca
      181 ggaactgctt accacagata tcctgtttgg cccatattct gctgtctctc tgttcctaac
      241 cttgatctga acttctctat tctcagttat gtattttcca tgccttgcaa aatggcgtta
      301 cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt ttctggagac
      361 taaataaaat cttttatttt atctatggct cgtactctat aggcttcagc tggtgatatt
      421 gttgagtcaa aactagagcc tggaccactg atatcctgtc tttaacaaat tggactaatc
      481 gcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
      541 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
      601 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
      661 ccttgcatgc accattcctt gcgccgtcgt tttacaacgt cgtgactggg aaaaccctgg
      721 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
      781 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct
      841 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct
      901 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
      961 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     1021 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     1081 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     1141 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1201 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1261 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1321 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1381 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1441 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1501 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1561 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1621 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1681 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     1741 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     1801 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     1861 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     1921 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     1981 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2041 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2101 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2161 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2221 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2281 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2341 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2401 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2461 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2521 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2581 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2641 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     2701 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     2761 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     2821 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     2881 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     2941 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3001 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3061 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3121 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     3181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     3241 tatgttgtgt ggaattgtga gcggataaca atttcacaat tctccccagc tctttctgcg
     3301 ctgtccataa tgaactattt caggagtatt ggagtatgtg gattcaaaac agtggctaat
     3361 tgagtcagga ttacttaata ttcgactcct accctcagaa tgttgttctc caacttcaag
     3421 aaactgttac ccatgaaaga cccccgctga cgggtagtca atcactcaga ggagaccctc
     3481 ccaaggaaca gcgagaccac aagtcggatg caactgcaag agggtttatt ggatacacgg
     3541 gtacccgggc gacgcagtct atcggaggac tggcgcgccg agtgaggggt tgtgggctct
     3601 tttattgagc tcggggagca gaagcgcgcg aacagaagcg agaagcgaac tgattggtta
     3661 gttcaaataa ggcacagggt catttcaggt ccttggggca ccctggaaac atctgatggt
     3721 tctctagctt gccaaaccta caggtggggt ctttcattcc cccctttttc tggagactaa
     3781 ataaaatctt ttattttatc gatgcatccc cgatccttat cgctatcgat tcacacaaaa
     3841 aaccaacaca cagatgtaat gaaaataaag atattttatt gcggccatcg tgatggcagg
     3901 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     3961 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     4021 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     4081 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     4141 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     4201 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     4261 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     4321 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     4381 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     4441 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     4501 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     4561 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     4621 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     4681 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgg
     4741 tggctctagc cttaagttcg agactgttgt gtcagaagaa tcaagctgat ctgagtccgg
     4801 acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     4861 ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc ggactgggtg ctcaggtagt
     4921 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     4981 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
     5041 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
     5101 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
     5161 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
     5221 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
     5281 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
     5341 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
     5401 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
     5461 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
     5521 tggcgaccgg taagctagct tgggctgcag gtcgaaaggc ccggagatga ggaagaggag
     5581 aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt
     5641 cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc accccttccc
     5701 agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct
     5761 acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact
     5821 tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg
     5881 ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg
     5941 gccggtggat gtggaatgtg tgcgaggcca gaggccactt gtgtagcgcc aagtgcccag
     6001 cggggctgct aaagcgcatg ctccagactg ccttgggaaa agcgcctccc ctacccggta
     6061 gaattcgaac gctgacgtca tcaacccgct ccaaggaatc gcgggcccag tgtcactagg
     6121 cgggaacacc cagcgcgcgt gcgccctggc aggaagatgg ctgtgaggga caggggagtg
     6181 gcgccctgca atatttgcat gtcgctatgt gttctgggaa atcaccataa acgtgaaatg
     6241 tctttggatt tgggaatctt ataagttccc tatcagtgat agagatcccc caggagccag
     6301 ctagatatta attcaagaga ttaatatcta gctggctcct gtttttaagc ttatcgatac
     6361 cgtcgacctc gagagatcga tctaattccg gcgcctagag aaggagtgag ggctggataa
     6421 agggaggatc gaggcggggt cgaacgagga ggttcaaggg ggagagacgg ggcggatgga
     6481 ggaagaggag gcggaggctt agggtgtaca aagggcttga cccagggagg ggggtcaaaa
     6541 gccaaggctt cccaggtcac gatgtagggg acctggtctg ggtgtccatg cgggccaggt
     6601 gaaaagacct tgatcttaac ctgggtgatg aggtctcggt taaaggtgcc gtctcgcggc
     6661 catccgacgt taaaggttgg ccattctgca gagcagaagg taacccaacg tctcttcttg
     6721 acatctaccg actggttgtg agcgatccgc tcgacatctt tccagtgacc taaggtcaaa
     6781 cttaagggag tggtaacagt ctggccctaa ttttcagaca aatacagaaa cacagtcaga
     6841 cagagacaac acagaacgat gctgcagcgc tgcagcagac aagacgcgcg gcttcggttc
     6901 caaaccgaaa gcaaaaattc agacggaggc gggaactgtt ttaggttctc gtctcctacc
     6961 agaaccacat atcctgacgg ggtcggattc cacatcgact cccttcctca ggtcgggcca
     7021 caaaaacggc ccccaaagtc cctgggacgt ctcccagggt tgcggccggg tgttcagaac
     7081 tcgtcagttc caccacgggt ccgccagata cagagctagt tagctaacta gtacagacgc
     7141 aggcgcaaac attagatgcc ggcacaaaca cgcacaaaga cagagacaga cacgaaacga
     7201 ccgctggcca gcttacctcc cggcgggggg gtcggtggtc cctgggcagg ggtctccaaa
     7261 tctcggtgga acctccaaat gaaagacccc cgaggtgggc agtcaatcaa tctgaggaga
     7321 ccctcccaag gatcagcgag tccacgattc ggatgcaaac agcaagaggc tttattggga
     7381 atacgggtac ccgggcgacg cagtctatcg gaggactggc gcgccgagtg aggggttgtg
     7441 ggctctttta ttg
//
