LOCUS       sequence-199427-        5233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 - SSX1 promoter drives self-inhibiting mKate2 expression 
            (Version 2 intron - see PMID: 29056342 for detailed information).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5233)
  AUTHORS   Nissim L, Wu MR, Pery E, Binder-Nissim A, Suzuki HI, Stupp D, 
            Wehrspaun C, Tabach Y, Sharp PA, Lu TK
  TITLE     Synthetic RNA-Based Immunomodulatory Gene Circuits for Cancer 
            Immunotherapy.
  JOURNAL   Cell. 2017 Nov 16;171(5):1138-1150.e15. doi: 
            10.1016/j.cell.2017.09.049. Epub 2017 Oct 19.
  PUBMED    29056342
REFERENCE   2  (bases 1 to 5233)
  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: 
            "10.1016/j.cell.2017.09.049"; journalName: "Cell"; date: 
            "2017-11-16- 16"; volume: "171"; issue: "5"; pages: "1138-1150.e15"
COMMENT     Sequence Label: pLN189 (SSX1p-[mK-Ex1]-[miR1-Mv2
            intron]-[mK-E...
FEATURES             Location/Qualifiers
     source          1..5233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..145
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     regulatory      1257..1266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     ncRNA           1568..1689
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1717..1731
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1788..1900
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(2731..2750)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2775..2822
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(3138..3155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3309..3897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3389..3408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4068..4859,4860..4928))
                     /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     4691..4710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4929..5033)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5060..5233,1..282)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5147..5166)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
       61 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      121 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      181 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      241 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttgccattcg ccattcaggc
      301 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagcccaagc
      361 taccatgata agtaagtaat attaaggtac gggaggtact tggaggtacc cggccgccct
      421 gcagggtagc cagatcatgg ctcactgcaa cctcgtactc ctgggctcaa gctatcctcc
      481 tacctcagcc tcctgagtaa cggactacag gcacaccacc ccacctcgct aattttattt
      541 attttttttg tagagaaaag agacagggta ttgctctgtt gcccagggtg gggtgcagtg
      601 gcatgatcat ggctcactgc aacctctgcc tcccaggttc aagtgatcct ccagctgtgg
      661 cctccctaag tgctgggatt acaaccgtga gccgccgcac cggcccaaat ttcttacgtc
      721 actacagagt tcctaggaaa aaatcccata cctgaaaaag atagaaactg acaggaagga
      781 tttgagatga tgacctgctt catatacact ccttattaaa actggataac aatgcaccac
      841 cgaggaggtg gggagggata ggaaaaatga aaagagaaaa tcagcgcatg cgtactctga
      901 tttgggaaga ctccaaagag aaaatcagag catgcgtact ctgaacttgg agtagccaat
      961 cccaggggat gctttaggcg ggaaagtcag agtttctgcc tccattttga gaaggttctg
     1021 tccctagagc ctagactgat agaccccaca tcagcttggc ttgtcccgcc tactgttctg
     1081 acttctgatt ggccagatgg agttcactaa ctgccctgat tggtccatca tcctggagca
     1141 atgacattgc agaatatttt ctcctcctcc agccacactt tgtcaccaac tgctgccaac
     1201 tcgccaccac tgctgccgac ctcgcaacca ctgctttgtc tctggcgcgc cagctcgcca
     1261 ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg
     1321 agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg
     1381 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     1441 acatcctggc taccagcttc atgtacggca gcaaaacctt catcaaccac acccagggca
     1501 tccccgactt ctttaagcag tccttccctg aggtaagtgt gctcgcttcg gcagcacata
     1561 tactatgttg aatgaggctt cagtacttta cagaatcgtt gcctgcacat cttggaaaca
     1621 cttgctggga ttacttcttc aggttaaccc aacagaaggc tcgagaaggt atattgctgt
     1681 tgacagtgag cgccgcttga agtctttaat taaatagtga agccacagat gtatttaatt
     1741 aaagacttca agcggtgcct actgcctcgg acttcaaggg gctagaattc aaggggctac
     1801 tttaggagca attatcttgt ttactaaaac tgaatacctt gctatctctt tgatacattt
     1861 ttacaaagct gaattaaaat ggtataaatt aaatcacttt tttcaattgt actaacttcg
     1921 agtcttcttt ttttttttca cagggcttca catgggagag agtcaccaca tacgaagacg
     1981 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
     2041 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
     2101 gctgggaggc ctccaccgag atgctgtacc ccgctgacgg cggcctggaa ggcagaagcg
     2161 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca
     2221 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac
     2281 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
     2341 ccagatactg cgacctccct agcaaactgg ggcacaaact taattgataa acataggatg
     2401 gcaagatcct ggtatcggtc tgcgagcacc gcttgaagtc tttaattaaa tacgcaccgc
     2461 ttgaagtctt taattaaata cgcaccgctt gaagtcttta attaaatacc tcgagcctct
     2521 ggccacatcg gttcctgcgg taagcgggac tctggggttc gaaatgaccg accaagcgac
     2581 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     2641 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     2701 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     2761 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     2821 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacgcca
     2881 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     2941 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggtatgtc
     3001 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     3061 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     3121 cagcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3181 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3241 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3301 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3361 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3421 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3481 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     3541 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3601 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3661 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3721 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     3781 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3841 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3901 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     3961 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4021 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4081 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4141 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4201 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4261 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4321 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4381 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4441 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     4501 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     4561 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     4621 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     4681 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     4741 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     4801 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     4861 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     4921 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     4981 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5041 ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     5101 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5161 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5221 cctttagggt tcc
//
