LOCUS       sequence-372398-        3858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PmPGK1-driven secreted Nluc 
            (A1-pA::A2-pA::PmPGK1-IgkSS-Nluc-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3858)
  AUTHORS   Haellman V, Strittmatter T, Bertschi A, Stucheli P, Fussenegger M
  TITLE     A versatile plasmid architecture for mammalian synthetic biology 
            (VAMSyB).
  JOURNAL   Metab Eng. 2021 Jul;66:41-50. doi: 10.1016/j.ymben.2021.04.003. Epub
            2021 Apr 18.
  PUBMED    33857582
REFERENCE   2  (bases 1 to 3858)
  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"; journalName: "Metab 
            Eng."; date: "2021-07"; volume: "66:41-50. doi"; pages: " 
            10.1016/j.ymben.2021.04.003. Epub 2021 Apr 18"
COMMENT     Sequence Label: pTS1037
FEATURES             Location/Qualifiers
     source          1..3858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(1..792,793..861))
                     /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     624..643
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(862..966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1127..1144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1133..1357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1503..1519)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1503..1519)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1559..2058
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1594..1617)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1991..2010
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     sig_peptide     2111..2167
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     CDS             2189..2701
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     protein_bind    2841..2862
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2979..2996)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3150..3738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3230..3249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
       61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
      121 cagtgctgca atgataccgc ggctcccacg ctcaccggct ccagatttat cagcaataaa
      181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
      241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
      301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
      361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
      421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
      481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
      541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
      601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
      661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
      721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
      781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
      841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
      901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccaaga tctccacgcg
     1021 tggtaccctc gaggagctct tcgaagcgga attcaccatg actagtgcta gcggatccac
     1081 cggtgtctag aaagctttga ggccggcctg caggcccgct gatcagcctc gactgtgcct
     1141 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1201 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1261 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1321 aatagcaggc atgctgggga tgcggtgggc tctatggtag cttaggaggc ggaaagaacc
     1381 agcgcgggag accgtcagtt ggtctccagc tatacaggtg caccgcttta taataaaact
     1441 gatacagaga acgttatcat gtgtgttggt tttttttgtg tgtgcggttt aagggagcct
     1501 cggtcatagc tgtttcctgt gtaaaacgac ggccagcgcg tggtaccctc gagctaccgg
     1561 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     1621 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     1681 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     1741 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     1801 ctcagtagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     1861 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     1921 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     1981 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     2041 tcatctccgg gcctttcgac ctccagcccg agctcggtat aggttcgaag cggaattcac
     2101 catgactagt gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc
     2161 cactggtgac gctagtggtg gttctggtat ggtcttcaca ctcgaagatt tcgttgggga
     2221 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
     2281 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
     2341 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca
     2401 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa
     2461 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta
     2521 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg
     2581 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct
     2641 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc
     2701 ggctagcgga tccaccggtg tctagaaagc tcagaaatta tgagagcgtt taataaagta
     2761 ttgactatgg ttgacaaaac gtgtgttgga gttttttgtg tgctttgtca tcaactgagc
     2821 cagtaggtcg agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     2881 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2941 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3001 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3061 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggattat caaaaaggat
     3781 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3841 gtaaacttgg tctgacag
//
