LOCUS       sequence-192978-        5452 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 5452)
  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 5452)
  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 delta1
FEATURES             Location/Qualifiers
     source          1..5452
                     /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    1476..2098
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1675..1694)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2122..2142)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2127..2456
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2307..2442
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2494..2513)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2495..2513)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2527..2543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2527..2543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2540..2562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2551..2567
                     /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(2575..2581,2582..2599,2600..2605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2758..2775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2929..3517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3009..3028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3688..4479,4480..4548))
                     /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     4311..4330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4549..4653)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4680..5135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4767..4786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4977..4998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5261..5283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5275..5292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5276..5292
                     /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 gatgggcagc gccagcccgg gtctgagcag cgtatccccc agccacctcc tgctgccccc
      781 cgacacggtg tcgcggacag gcttggagaa ggcggcagcg ggggcagtgg gtctcgagag
      841 acgggactgg agtcccagtc cacccgccac gcccgagcag ggcctgtccg ccttctacct
      901 ctcctacttt gacatgctgt accctgagga cagcagctgg gcagccaagg cccctggggc
      961 cagcagtcgg gaggagccac ctgaggagcc tgagcagtgc ccggtcattg acagccaagc
     1021 cccagcgggc agcctggact tggtgcccgg cgggctgacc ttggaggagc actcgctgga
     1081 gcaggtgcag tccatggtgg tgggcgaagt gctcaaggac atcgagacgg cctgcaagct
     1141 gctcaacatc accgcagatc ccatggactg gagccccagc aatgtgcaga agtggctcct
     1201 gtggacagag caccaatacc ggctgccccc catgggcaag gccttccagg agctggcggg
     1261 taaggagctg tgcgccatgt cggaggagca gttccgccag cgctcgcccc tgggtgggga
     1321 tgtgctgcac gcccacctgg acatctggaa gtcagcggcc tggatgaaag agcggacttc
     1381 acctggggcg attcactact gtgcctcgac cagtgaggag agctggaccg acagcgaggt
     1441 ggactcatca tgctccgggc agccctgagg atcccgggtg gcatccctgt gacccctccc
     1501 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     1561 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     1621 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     1681 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     1741 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     1801 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     1861 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     1921 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg
     1981 aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga catttgagtt
     2041 gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg
     2101 gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2161 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2221 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2281 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2341 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2401 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2461 tcctcgagga actgaaaaac cagaaagtta attccctata gtgagtcgta ttaaattcgt
     2521 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2581 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2641 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2701 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2761 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2821 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2881 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2941 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3001 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3061 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3121 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3181 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3241 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3301 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3361 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3421 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3481 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3541 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3601 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3661 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3721 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3781 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3841 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3901 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3961 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     4021 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     4081 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     4141 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4201 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4261 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4321 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4381 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4441 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4501 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4561 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4621 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4681 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4741 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4801 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4861 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4921 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4981 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5041 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5101 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     5161 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     5221 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     5281 acgacggcca gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta
     5341 gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta
     5401 tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg tt
//
