LOCUS       sequence-386330-        7678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral Tet response vector containing GFP tagged Crumbs3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7678)
  AUTHORS   Schluter MA, Pfarr CS, Pieczynski J, Whiteman EL, Hurd TW, Fan S, 
            Liu CJ, Margolis B
  TITLE     Trafficking of Crumbs3 during cytokinesis is crucial for lumen 
            formation.
  JOURNAL   Mol Biol Cell. 2009 Nov;20(22):4652-63. doi: 
            10.1091/mbc.e09-02-0137. Epub 2009 Sep 23.
  PUBMED    19776356
REFERENCE   2  (bases 1 to 7678)
  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"; doi: 
            "10.1091/mbc.e09-02-0137"; journalName: "Mol Biol Cell"; date: 
            "2009-11"; volume: "20"; issue: "22"; pages: "4652-63"
COMMENT     Sequence Label: pRevTRE-hygro-GFP-Crumbs3a WT
FEATURES             Location/Qualifiers
     source          1..7678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     578..597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(816..920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     988..1006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     LTR             1578..1753
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    1816..2015
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2216..2632
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2562..2584
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2600..2616
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             2688..3707
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     CDS             2688..3707
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     protein_bind    join(4011..4029,4030..4052,4053..4071,4072..4094,4095..4113,
                     4114..4136,4137..4155,4156..4178,4179..4197,4198..4220,
                     4221..4239,4240..4262,4263..4281)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(4053..4071,4072..4094,4095..4113,4114..4136,4137..4155,
                     4156..4178,4179..4197,4198..4220,4221..4239,4240..4262,
                     4263..4281,4282..4304,4305..4323)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(4095..4113,4114..4136,4137..4155,4156..4178,4179..4197,
                     4198..4220,4221..4239,4240..4262,4263..4281,4282..4304,
                     4305..4323,4324..4346,4347..4365)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     protein_bind    join(4137..4155,4156..4178,4179..4197,4198..4220,4221..4239,
                     4240..4262,4263..4281,4282..4304,4305..4323,4324..4346,
                     4347..4365,4366..4388,4389..4407)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        4440..4478
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4475..4499
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             join(4757..4759,4760..5470)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLKE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4799..4820)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5060..5079)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     LTR             5744..6337
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6440..6462)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6548..6688
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6603..6622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6703..6720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6874..7462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6954..6973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7633..7678,1..746,747..815))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
       61 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      121 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      181 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      241 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
      301 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      361 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
      421 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
      481 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
      541 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
      601 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
      661 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
      721 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
      781 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
      841 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
      901 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
      961 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
     1021 gaattcatac cagatcaccg aaaactgtcc tccaaatgtg tccccctcac actcccaaat
     1081 tcgcgggctt ctgcctctta gaccactcta ccctattccc cacactcacc ggagccaaag
     1141 ccgcggccct tccgtttctt tgcttttgaa agaccccacc cgtaggtggc aagctagctt
     1201 aagtaacgcc actttgcaag gcatggaaaa atacataact gagaatagaa aagttcagat
     1261 caaggtcagg aacaaagaaa cagctgaata ccaaacagga tatctgtggt aagcggttcc
     1321 tgccccggct cagggccaag aacagatgag acagctgagt gatgggccaa acaggatatc
     1381 tgtggtaagc agttcctgcc ccggctcggg gccaagaaca gatggtcccc agatgcggtc
     1441 cagccctcag cagtttctag tgaatcatca gatgtttcca gggtgcccca aggacctgaa
     1501 aatgaccctg taccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     1561 cttccgctct ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcttc
     1621 cgatagactg cgtcgcccgg gtacccgtat tcccaataaa gcctcttgct gtttgcatcc
     1681 gaatcgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccacgacg
     1741 ggggtctttc atttgggggc tcgtccggga tttggagacc cctgcccagg gaccaccgac
     1801 ccaccaccgg gaggtaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
     1861 tatgtttgat gttatgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     1921 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     1981 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     2041 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     2101 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     2161 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     2221 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     2281 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     2341 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     2401 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     2461 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     2521 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     2581 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattcc
     2641 gatctgatag cttgccacaa cccgtaccaa agatggatag atccggaaag cctgaactca
     2701 ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc
     2761 agctctcgga gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg
     2821 tcctgcgggt aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact
     2881 ttgcatcggc cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc
     2941 tgacctattg catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg
     3001 aactgcccgc tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc
     3061 ttagccagac gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat
     3121 ggcgtgattt catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg
     3181 acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg
     3241 actgccccga agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg
     3301 acaatggccg cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat
     3361 acgaggtcgc caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc
     3421 gctacttcga gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc
     3481 tccgcattgg tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag
     3541 cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta
     3601 cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg
     3661 atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa ggaatagagt agatgccgac
     3721 cgaacaagag ctgatttcga gaacgcctca gccagcaact cgcgcgagcc tagcaaggca
     3781 aatgcgagag aacggcctta cgcttggtgg cacagttctc gtccacagtt cgctaagctc
     3841 gctcggctgg gtcgcgggag ggccggtcgc agtgattcag gcccttctgg attgtgttgg
     3901 tccccagggc acgattgtca tgcccacgca ctcgggtgat ctgactgatc ccgcagattg
     3961 gagatcgccg cccgtgcctg ccgattgggt gcagatctcg agtttaccac tccctatcag
     4021 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     4081 gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
     4141 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
     4201 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     4261 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     4321 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     4381 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agctcggtac ccgggtcgag
     4441 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
     4501 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
     4561 tccgcggccc cgaattcgag ctcggtaccc gggatccatg gcgaaccccg ggctggggct
     4621 gcttctggcg ctgggcctgc cgttcctgct ggcccgctgg ggccgagcct gggggcaaat
     4681 acagaccact tctgcaaatg agaatagcac tgttttgcct tcatccacca gctccagctc
     4741 cgacggaggt ggaggggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     4801 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     4861 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
     4921 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
     4981 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     5041 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     5101 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     5161 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     5221 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     5281 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     5341 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     5401 cgatcacatg gtccttaagg agttcgtgac cgccgccggg atcactctcg gcatggacga
     5461 gctgtacaag ggtggaggcg atggcaacct gcgtccagaa gccatcactg ctatcatcgt
     5521 ggtcttctcc ctcttggctg ccttgctcct ggctgtgggg ctggcactgt tggtgcggaa
     5581 gcttcgggag aagcggcaga cggagggcac ctaccggccc agtagcgagg agcaggtggg
     5641 tgcccgcgta ccaccgaccc ccaacctcaa gttgccgccg gaagagcggc tcatctgaat
     5701 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     5761 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     5821 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     5881 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     5941 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     6001 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     6061 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     6121 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     6181 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     6241 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     6301 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
     6361 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc ctcgcgcgtt
     6421 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6481 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6541 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     6601 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6661 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     6721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtcta
//
