LOCUS       sequence-192982-        4984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human SPDEF truncation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4984)
  AUTHORS   Lo YH, Noah TK, Chen MS, Zou W, Borras E, Vilar E, Shroyer NF
  TITLE     SPDEF Induces Quiescence of Colorectal Cancer Cells by Changing the 
            Transcriptional Targets of beta-catenin.
  JOURNAL   Gastroenterology. 2017 Jul;153(1):205-218.e8. doi: 
            10.1053/j.gastro.2017.03.048. Epub 2017 Apr 5.
  PUBMED    28390865
REFERENCE   2  (bases 1 to 4984)
  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: 
            "10.1053/j.gastro.2017.03.048"; journalName: "Gastroenterology"; 
            date: "2017-07"; volume: "153"; issue: "1"; pages: "205-218.e8"
COMMENT     Sequence Label: pCMV7.1-Flag-NLS-hSPDEF delta3
FEATURES             Location/Qualifiers
     source          1..4984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             614..679
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             701..721
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1008..1630
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1207..1226)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(1654..1674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1659..1988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1839..1974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2026..2045)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2027..2045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2059..2075)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2059..2075)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2072..2094)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2083..2099
                     /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(2107..2113,2114..2131,2132..2137))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2152..2173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2290..2307)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2461..3049)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2541..2560)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3220..4011,4012..4080))
                     /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     3843..3862
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4081..4185)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4212..4667
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4299..4318)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4509..4530
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4793..4815
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4807..4824
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4808..4824
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattaacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
      661 caaggatgac gatgacaagc ttgcggccgc gaattccgcc cccaagaaaa agagaaaggt
      721 gtgctccggg cagcccatcc acctgtggca gttcctcaag gagttgctac tcaagcccca
      781 cagctatggc cgcttcatta ggtggctcaa caaggagaag ggcatcttca aaattgagga
      841 ctcagcccag gtggcccggc tgtggggcat ccgcaagaac cgtcccgcca tgaactacga
      901 caagctgagc cgctccatcc gccagtatta caagaagggc atcatccgga agccagacat
      961 ctcccagcgc ctcgtctacc agttcgtgca ccccatctga ggatcccggg tggcatccct
     1021 gtgacccctc cccagtgcct ctcctggccc tggaagttgc cactccagtg cccaccagcc
     1081 ttgtcctaat aaaattaagt tgcatcattt tgtctgacta ggtgtccttc tataatatta
     1141 tggggtggag gggggtggta tggagcaagg ggcaagttgg gaagacaacc tgtagggcct
     1201 gcggggtcta ttgggaacca agctggagtg cagtggcaca atcttggctc actgcaatct
     1261 ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc cgagttgttg ggattccagg
     1321 catgcatgac caggctcagc taatttttgt ttttttggta gagacggggt ttcaccatat
     1381 tggccaggct ggtctccaac tcctaatctc aggtgatcta cccaccttgg cctcccaaat
     1441 tgctgggatt acaggcgtga accactgctc ccttccctgt ccttctgatt ttaaaataac
     1501 tataccagca ggaggacgtc cagacacagc ataggctacc tggccatgcc caaccggtgg
     1561 gacatttgag ttgcttgctt ggcactgtcc tctcatgcgt tgggtccact cagtagatgc
     1621 ctgttgaatt gggtacgcgg ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa
     1681 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     1741 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     1801 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1861 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1921 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1981 tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt taattcccta tagtgagtcg
     2041 tattaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2101 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     2161 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2221 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2281 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2341 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2401 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2461 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2521 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2581 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2641 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2701 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2761 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2821 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2881 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2941 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3001 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3061 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3121 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3181 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3241 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3301 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3361 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3421 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3481 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3541 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3601 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3661 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3721 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3781 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3841 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3901 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3961 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4021 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4081 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4141 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4201 agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4261 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4321 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4381 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4441 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4501 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4561 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4621 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
     4681 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4741 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4801 cgacgttgta aaacgacggc cagtgccaag ctgatctata cattgaatca atattggcaa
     4861 ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     4921 acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat atgaccgcca
     4981 tgtt
//
