LOCUS       sequence-365605-        3858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PmPGK1-driven secreted Nluc 
            (A1-pA::PmPGK1-IgkSS-Nluc-pA::A3-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: pTS1036
FEATURES             Location/Qualifiers
     source          1..3858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(2..793,794..862))
                     /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     625..644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(863..967)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1128..1145)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1134..1358
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1406..1905
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1441..1464)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1838..1857
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     sig_peptide     1958..2014
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     CDS             2036..2548
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(2670..2686)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2670..2686)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    2842..2863
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2980..2997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3151..3739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3231..3250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
       61 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
      121 ccagtgctgc aatgataccg cggctcccac gctcaccggc tccagattta tcagcaataa
      181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
      241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
      481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
      541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
      601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      961 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccaag atctccacgc
     1021 gtggtaccct cgaggagctc ttcgaagcgg aattcaccat gactagtgct agcggatcca
     1081 ccggtgtcta gaaagctttg aggccggcct gcaggcccgc tgatcagcct cgactgtgcc
     1141 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1201 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1261 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     1321 caatagcagg catgctgggg atgcggtggg ctctatggta gcttaggagg cggaaagaac
     1381 cagcgcgtgg taccctcgag ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     1441 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     1501 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     1561 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     1621 aggacgtgac aaatggaagt agcacgtctc agtagtctcg tgcagatgga cagcaccgct
     1681 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     1741 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     1801 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     1861 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctc cagcccgagc
     1921 tcggtatagg ttcgaagcgg aattcaccat gactagtgag acagacacac tcctgctatg
     1981 ggtactgctg ctctgggttc caggttccac tggtgacgct agtggtggtt ctggtatggt
     2041 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
     2101 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
     2161 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
     2221 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
     2281 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
     2341 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
     2401 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
     2461 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
     2521 ctggcggctg tgcgaacgca ttctggcggc tagcggatcc accggtgtct agaaagctat
     2581 acaggtgcac cgctttataa taaaactgat acagagaacg ttatcatgtg tgttggtttt
     2641 ttttgtgtgt gcggtttaag ggagcctcgg tcatagctgt ttcctgtgta aaacgacggc
     2701 cagcgcggga gacggtcagt tcgtctccag ctcagaaatt atgagagcgt ttaataaagt
     2761 attgactatg gttgacaaaa cgtgtgttgg agttttttgt gtgctttgtc atcaactgag
     2821 ccagtaggtc gagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2881 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     2941 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     3001 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3061 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3121 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3181 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3241 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3301 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3361 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3421 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3481 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3541 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     3601 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3661 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggatta tcaaaaagga
     3781 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     3841 agtaaacttg gtctgaca
//
