LOCUS       sequence-364216-        5220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-controlled FURIN protease recognition 
            site (PhCMV-FURINsite-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5220)
  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 5220)
  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: pTS2409-Tier1-PhCMV-FURINsite
FEATURES             Location/Qualifiers
     source          1..5220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        435..638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     589..609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     683..702
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        683..701
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(826..843)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    832..1056
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1102..1530
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1189..1208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1399..1420
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1539..1559)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1544..1873
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1724..1859
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1786..1805
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1940..2734
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1994..2013)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2604..2623
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2908..3029
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2945..2964)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2999..3018
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3078..3094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3078..3094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3091..3113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3102..3118
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3126..3132,3133..3150,3151..3156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3171..3192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4239..5030,5031..5099))
                     /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     4862..4881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5100..5204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgtcgacgg atcgggagat ctccacgcgt ggtaccctcg agttgacatt
       61 gattattgag tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      121 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      181 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      241 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      301 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      361 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      421 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      481 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      541 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      601 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      661 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctgttcgaa
      721 gcggaattca ccatgactag tggtggttct ggtagagcca gatacaagag agctagcgga
      781 tccaccggtg tctagaaagc tttgaggccg gcctgcaggg atcagcctcg actgtgcctt
      841 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
      901 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
      961 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     1021 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1081 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1141 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1201 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     1261 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     1321 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     1381 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     1441 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     1501 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     1561 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     1621 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     1681 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     1741 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     1801 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     1861 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     1921 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     1981 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     2041 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     2101 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     2161 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     2221 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     2281 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     2341 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     2401 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     2461 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     2521 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     2581 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     2641 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     2701 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     2761 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     2821 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     2881 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     2941 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3001 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     3061 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3241 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
//
