LOCUS       sequence-201286-        5216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module 1 - SSX1 promoter drives self-inhibiting mKate2 expression 
            (Version 1 intron - see PMID: 29056342 for detailed information).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5216)
  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 5216)
  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: pLN520 (SSX1p-[mK-Ex1]-[miR1-Mv1
            intron]-[mK-E...
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1081..1090
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     ncRNA           1392..1485
                     /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           1532..1546
                     /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           1574..1702
                     /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(2533..2552)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2577..2624
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(2945..2962)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3116..3704)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3196..3215)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3875..4666,4667..4735))
                     /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     4498..4517
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4736..4840)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4867..5216,1..106)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4954..4973)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5164..5185
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
       61 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttgcca ttcgccattc
      121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagccc
      181 aagctaccat gataagtaag taatattaag gtacgggagg tacttggagg tacccggccg
      241 ccctgcaggg tagccagatc atggctcact gcaacctcgt actcctgggc tcaagctatc
      301 ctcctacctc agcctcctga gtaacggact acaggcacac caccccacct cgctaatttt
      361 atttattttt tttgtagaga aaagagacag ggtattgctc tgttgcccag ggtggggtgc
      421 agtggcatga tcatggctca ctgcaacctc tgcctcccag gttcaagtga tcctccagct
      481 gtggcctccc taagtgctgg gattacaacc gtgagccgcc gcaccggccc aaatttctta
      541 cgtcactaca gagttcctag gaaaaaatcc catacctgaa aaagatagaa actgacagga
      601 aggatttgag atgatgacct gcttcatata cactccttat taaaactgga taacaatgca
      661 ccaccgagga ggtggggagg gataggaaaa atgaaaagag aaaatcagcg catgcgtact
      721 ctgatttggg aagactccaa agagaaaatc agagcatgcg tactctgaac ttggagtagc
      781 caatcccagg ggatgcttta ggcgggaaag tcagagtttc tgcctccatt ttgagaaggt
      841 tctgtcccta gagcctagac tgatagaccc cacatcagct tggcttgtcc cgcctactgt
      901 tctgacttct gattggccag atggagttca ctaactgccc tgattggtcc atcatcctgg
      961 agcaatgaca ttgcagaata ttttctcctc ctccagccac actttgtcac caactgctgc
     1021 caactcgcca ccactgctgc cgacctcgca accactgctt tgtctctggc gcgccagctc
     1081 gccaccatgg tgtctaaggg cgaagagctg attaaggaga acatgcacat gaagctgtac
     1141 atggagggca ccgtgaacaa ccaccacttc aagtgcacat ccgagggcga aggcaagccc
     1201 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     1261 ttcgacatcc tggctaccag cttcatgtac ggcagcaaaa ccttcatcaa ccacacccag
     1321 ggcatccccg acttctttaa gcagtccttc cctgaggtaa gtgtgctcgc ttcggcagca
     1381 catatactat gttgaatgag gcttcagtac tttacagaat cgttgcctgc acatcttgga
     1441 aacacttgct gggattactt cttcaggtta acccaacaga aggctcgagt gctgttgaca
     1501 gtgagcgccg cttgaagtct ttaattaaat agtgaagcca cagatgtatt taattaaaga
     1561 cttcaagcgg tgcctactgc ctcggagaat tcaaggggct actttaggag caattatctt
     1621 gtttactaaa actgaatacc ttgctatctc tttgatacat ttttacaaag ctgaattaaa
     1681 atggtataaa ttaaatcact tttttcaatt gtactaactt cgagtcttct tttttttttt
     1741 cacagggctt cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta
     1801 cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga
     1861 acttcccatc caacggccct gtgatgcaga agaaaacact cggctgggag gcctccaccg
     1921 agatgctgta ccccgctgac ggcggcctgg aaggcagaag cgacatggcc ctgaagctcg
     1981 tgggcggggg ccacctgatc tgcaacttga agaccacata cagatccaag aaacccgcta
     2041 agaacctcaa gatgcccggc gtctactatg tggacagaag actggaaaga atcaaggagg
     2101 ccgacaaaga gacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc
     2161 ctagcaaact ggggcacaaa cttaattgat aaacatagga tggcaagatc ctggtatcgg
     2221 tctgcgagca ccgcttgaag tctttaatta aatacgcacc gcttgaagtc tttaattaaa
     2281 tacgcaccgc ttgaagtctt taattaaata cctcgagcct ctggccacat cggttcctgc
     2341 ggtaagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     2401 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     2461 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     2521 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     2581 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     2641 gtcccagggc tggcactctg tcgatacccc accgagacgc cattggggcc aatacgcccg
     2701 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     2761 caacgtcggg gcggcaggcc ctgccatagc ctcagctcga gtatgtcgac cgatgccctt
     2821 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     2881 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctcttccg
     2941 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3001 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3061 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3121 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3181 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3241 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3301 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3361 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3421 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3481 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3541 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3601 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3661 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3721 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3781 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3841 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3901 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3961 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4021 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4081 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4141 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4201 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4261 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4321 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4381 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4441 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4501 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4561 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4621 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4681 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4741 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4801 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4861 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4921 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4981 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     5041 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     5101 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     5161 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattct
//
