LOCUS       sequence-365611-        5919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-2 vector encoding PhCMV-driven SEAP coupled to fluorescent 
            reporter iRFP670 (PhCMV-SEAP-p2A-iRFP670-pA::A2-pA::A3-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  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 5919)
  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: pTS1039
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(493..1284,1285..1353))
                     /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     1116..1135
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1354..1458)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1528..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2062..2082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2156..2175
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2156..2174
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3793..3849
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3856..4785
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="ARKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAGA
                     FFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLHR
                     HDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVML
                     YRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSRI
                     VPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYEP
                     RAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     primer_bind     complement(4846..4863)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4852..5076
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5222..5238)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5222..5238)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5394..5415
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5532..5549)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5703..5919,1..372))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5783..5802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
       61 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      121 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      181 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      241 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
      301 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
      361 aaaggatctc aagaagatcc tttgatcttt tctacgggga ttatcaaaaa ggatcttcac
      421 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
      481 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
      541 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
      601 accatctggc cccagtgctg caatgatacc gcggctccca cgctcaccgg ctccagattt
      661 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
      721 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
      781 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
      841 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
      901 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
      961 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1021 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1081 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1141 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1201 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1261 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     1321 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     1381 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     1441 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccaa
     1501 gatctccacg cgtggtaccc tcgagttgac attgattatt gagtagttat taatagtaat
     1561 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
     2161 cgactcacta tagggagacc caagctgttc gaagcggaat tcaccatgac tagtctgctg
     2221 ctgctgctgc tgggcctgag actgcagctg agcctgggca tcatccccgt ggaggaggag
     2281 aaccccgact tctggaacag agaggccgcc gaggccctgg gcgccgccaa gaagctgcag
     2341 cccgcccaga ccgccgccaa gaacctgatc atcttcctgg gcgacggcat gggcgtgagc
     2401 accgtgaccg ccgccagaat cctgaagggc cagaagaagg acaagctggg ccccgagatc
     2461 cccctggcca tggacagatt cccctacgtg gccctgagca agacctacaa cgtggacaag
     2521 cacgtgcccg acagcggcgc caccgccacc gcctacctgt gcggcgtgaa gggcaacttc
     2581 cagaccatcg gcctgagcgc cgccgccaga ttcaaccagt gcaacaccac cagaggcaac
     2641 gaggtgatca gcgtgatgaa cagagccaag aaggccggca agagcgtggg cgtggtgacc
     2701 accaccagag tgcagcacgc cagccccgcc ggcacctacg cccacaccgt gaacagaaac
     2761 tggtacagcg acgccgacgt gcccgccagc gccagacagg agggctgcca ggacatcgcc
     2821 acccagctga tcagcaacat ggacatcgac gtgatcctgg gcggcggcag aaagtacatg
     2881 ttcagaatgg gcacccccga ccccgagtac cccgacgact acagccaggg cggcaccaga
     2941 ctggacggca agaacctggt gcaggagtgg ctggccaaga gacagggcgc cagatacgtg
     3001 tggaacagaa ccgagctgat gcaggccagc ctggacccca gcgtgaccca cctgatgggc
     3061 ctgttcgagc ccggcgacat gaagtacgag atccacagag acagcaccct ggaccccagc
     3121 ctgatggaga tgaccgaggc cgccctgaga ctgctgagca gaaaccccag aggcttcttc
     3181 ctgttcgtgg agggcggcag aatcgaccac ggccaccacg agagcagagc ctacagagcc
     3241 ctgaccgaga ccatcatgtt cgacgacgcc atcgagagag ccggccagct gaccagcgag
     3301 gaggacaccc tgagcctggt gaccgccgac cacagccacg tgttcagctt cggcggctac
     3361 cccctgagag gcagcagcat cttcggcctg gcccccggca aggccagaga cagaaaggcc
     3421 tacaccgtgc tgctgtacgg caacggcccc ggctacgtgc tgaaggacgg cgccagaccc
     3481 gacgtgaccg agagcgagag cggcagcccc gagtacagac agcagagcgc cgtgcccctg
     3541 gacgaggaga cccacgccgg cgaggacgtg gccgtgttcg ccagaggccc ccaggcccac
     3601 ctggtgcacg gcgtgcagga gcagaccttc atcgcccacg tgatggcctt cgccgcctgc
     3661 ctggagccct acaccgcctg cgacctggcc ccccccgccg gcaccaccga cgccgcccac
     3721 cccggctaca gcagagtggg cgccgccggc agattcgagc agaccgctag tggtggttct
     3781 ggtggaagcg gagctactaa cttcagcctg ctgaagcagg ctggtgacgt ggaggagaac
     3841 cctggacctg ctagtgcgcg taaggtcgat ctcacctcct gcgatcgcga gccgatccac
     3901 atccccggca gcattcagcc gtgcggctgc ctgcttgcct gcgacgcgca ggcggtgcgg
     3961 atcacgcgca ttacggaaaa tgccggcgcg ttctttggac gcgaaactcc gcgggtcggt
     4021 gagctactcg ccgattactt cggcgagacc gaagcccatg cgctgcgcaa cgcactggcg
     4081 cagtcctccg atccaaagcg accggcgctg atcttcggtt ggcgcgacgg cctgaccggc
     4141 cgcaccttcg acatctcact gcatcgccat gacggtacat cgatcatcga gttcgagcct
     4201 gcggcggccg aacaggccga caatccgctg cggctgacgc ggcagatcat cgcgcgcacc
     4261 aaagaactga agtcgctcga agagatggcc gcacgggtgc cgcgctatct gcaggcgatg
     4321 ctcggctatc accgcgtgat gttgtaccgc ttcgcggacg acggctccgg gatggtgatc
     4381 ggcgaggcga agcgcagcga cctcgagagc tttctcggtc agcactttcc ggcgtcgctg
     4441 gtcccgcagc aggcgcggct actgtacttg aagaacgcga tccgcgtggt ctcggattcg
     4501 cgcggcatca gcagccggat cgtgcccgag cacgacgcct ccggcgccgc gctcgatctg
     4561 tcgttcgcgc acctgcgcag catctcgccc tgccatctcg aatttctgcg gaacatgggc
     4621 gtcagcgcct cgatgtcgct gtcgatcatc attgacggca cgctatgggg attgatcatc
     4681 tgtcatcatt acgagccgcg tgccgtgccg atggcgcagc gcgtcgcggc cgaaatgttc
     4741 gccgacttct tatcgctgca cttcaccgcc gcccaccacc aacgcgctag cggatccacc
     4801 ggtgtctaga aagctttgag gccggcctgc aggcccgctg atcagcctcg actgtgcctt
     4861 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4921 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4981 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5041 atagcaggca tgctggggat gcggtgggct ctatggtagc ttaggaggcg gaaagaacca
     5101 gcgcgggaga ccgtcagttg gtctccagct atacaggtgc accgctttat aataaaactg
     5161 atacagagaa cgttatcatg tgtgttggtt ttttttgtgt gtgcggttta agggagcctc
     5221 ggtcatagct gtttcctgtg taaaacgacg gccagcgcgg gagacggtca gttcgtctcc
     5281 agctcagaaa ttatgagagc gtttaataaa gtattgacta tggttgacaa aacgtgtgtt
     5341 ggagtttttt gtgtgctttg tcatcaactg agccagtagg tcgagggtgc ctaatgagtg
     5401 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     5461 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5521 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5581 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5881 agcgtggcgc tttctcatag ctcacgctgt aggtatctc
//
