LOCUS       sequence-365623-        3807 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PmPGK1-driven secreted Nluc 
            (PmPGK1-IgkSS-Nluc-pA::A2-pA::A3-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3807)
  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 3807)
  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: pTS1035
FEATURES             Location/Qualifiers
     source          1..3807
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(31..822,823..891))
                     /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     654..673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(892..996)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1069..1568
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1104..1127)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1501..1520
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     sig_peptide     1621..1677
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     CDS             1699..2211
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2272..2289)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2278..2502
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2648..2664)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2648..2664)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2820..2841
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2958..2975)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3129..3717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3209..3228)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
       61 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
      121 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc ggctcccacg
      181 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
      241 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
      301 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
      361 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
      421 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
      481 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
      541 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
      601 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
      661 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
      721 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
      781 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
      841 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
      901 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      961 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     1021 tgacgtctaa gaaaccaaga tctccacgcg tggtaccctc gagctaccgg gtaggggagg
     1081 cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct
     1141 acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc
     1201 cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc
     1261 ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcagtagtc
     1321 tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc
     1381 aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg
     1441 gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg
     1501 cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg
     1561 gcctttcgac ctccagcccg agctcggtat aggttcgaag cggaattcac catgactagt
     1621 gagacagaca cactcctgct atgggtactg ctgctctggg ttccaggttc cactggtgac
     1681 gctagtggtg gttctggtat ggtcttcaca ctcgaagatt tcgttgggga ctggcgacag
     1741 acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag tttgtttcag
     1801 aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga aaatgggctg
     1861 aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca aatgggccag
     1921 atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg
     1981 cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg
     2041 ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg
     2101 aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga
     2161 gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc ggctagcgga
     2221 tccaccggtg tctagaaagc tttgaggccg gcctgcaggc ccgctgatca gcctcgactg
     2281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggtagcttag gaggcggaaa
     2521 gaaccagcgc gggagaccgt cagttggtct ccagctatac aggtgcaccg ctttataata
     2581 aaactgatac agagaacgtt atcatgtgtg ttggtttttt ttgtgtgtgc ggtttaaggg
     2641 agcctcggtc atagctgttt cctgtgtaaa acgacggcca gcgcgggaga cggtcagttc
     2701 gtctccagct cagaaattat gagagcgttt aataaagtat tgactatggt tgacaaaacg
     2761 tgtgttggag ttttttgtgt gctttgtcat caactgagcc agtaggtcga gggtgcctaa
     2821 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     2881 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     2941 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3001 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3061 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3121 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3181 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3241 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     3301 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     3361 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     3421 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     3481 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     3541 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     3601 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     3661 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     3721 gatcctttga tcttttctac ggggattatc aaaaaggatc ttcacctaga tccttttaaa
     3781 ttaaaaatga agttttaaat caatcta
//
