LOCUS       sequence-365618-        3939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PhCMV-driven secreted Nluc 
            (PhCMV-IgkSS-Nluc-pA::A2-pA::A3-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3939)
  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 3939)
  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: pTS1031
FEATURES             Location/Qualifiers
     source          1..3939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(891..1682,1683..1751))
                     /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     1514..1533
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1752..1856)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1926..2305
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2306..2509
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2460..2480
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2554..2573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2554..2572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     2613..2669
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     CDS             2691..3203
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(3264..3281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3270..3494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3640..3656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3640..3656)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    3812..3833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      841 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      901 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      961 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1021 atgataccgc ggctcccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1081 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1141 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1201 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1261 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1321 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1381 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1441 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1501 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1561 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1621 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1681 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1741 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1801 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1861 tttccccgaa aagtgccacc tgacgtctaa gaaaccaaga tctccacgcg tggtaccctc
     1921 gagttgacat tgattattga gtagttatta atagtaatca attacggggt cattagttca
     1981 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2041 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2101 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2161 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2221 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2281 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2341 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2401 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2461 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
     2521 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
     2581 agctgttcga agcggaattc accatgacta gtgagacaga cacactcctg ctatgggtac
     2641 tgctgctctg ggttccaggt tccactggtg acgctagtgg tggttctggt atggtcttca
     2701 cactcgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     2761 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     2821 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     2881 atgaaggtct gagcggcgac caaatgggcc agatcgaaaa aatttttaag gtggtgtacc
     2941 ctgtggatga tcatcacttt aaggtgatcc tgcactatgg cacactggta atcgacgggg
     3001 ttacgccgaa catgatcgac tatttcggac ggccgtatga aggcatcgcc gtgttcgacg
     3061 gcaaaaagat cactgtaaca gggaccctgt ggaacggcaa caaaattatc gacgagcgcc
     3121 tgatcaaccc cgacggctcc ctgctgttcc gagtaaccat caacggagtg accggctggc
     3181 ggctgtgcga acgcattctg gcggctagcg gatccaccgg tgtctagaaa gctttgaggc
     3241 cggcctgcag gcccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3301 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3361 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3421 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3481 ggtgggctct atggtagctt aggaggcgga aagaaccagc gcgggagacc gtcagttggt
     3541 ctccagctat acaggtgcac cgctttataa taaaactgat acagagaacg ttatcatgtg
     3601 tgttggtttt ttttgtgtgt gcggtttaag ggagcctcgg tcatagctgt ttcctgtgta
     3661 aaacgacggc cagcgcggga gacggtcagt tcgtctccag ctcagaaatt atgagagcgt
     3721 ttaataaagt attgactatg gttgacaaaa cgtgtgttgg agttttttgt gtgctttgtc
     3781 atcaactgag ccagtaggtc gagggtgcct aatgagtgag ctaactcaca ttaattgcgt
     3841 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3901 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgct
//
