LOCUS       sequence-364240-        5223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-controlled tabacco etch virus protease 
            recognition site (PhCMV-TEVsite-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5223)
  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 5223)
  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: pTS2408-Tier1-PhCMV-TEVsite
FEATURES             Location/Qualifiers
     source          1..5223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     727..746
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        727..745
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             798..818
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(873..890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    879..1103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1149..1577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1236..1255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1446..1467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1586..1606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1591..1920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1771..1906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1833..1852
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1987..2781
                     /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(2041..2060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2651..2670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2955..3076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2992..3011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3046..3065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3125..3141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3125..3141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3138..3160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3149..3165
                     /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(3173..3179,3180..3197,3198..3203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3218..3239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3356..3373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3527..4115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3607..3626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4286..5077,5078..5146))
                     /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     4909..4928
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5147..5223,1..28))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
       61 acggatcggg agatctccac gcgtggtacc ctcgagttga cattgattat tgagtagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
      721 cgaaattaat acgactcact atagggagac ccaagctgtt cgaagcggaa ttcaccatga
      781 ctagtggtgg ttctggtgag aatctgtact ttcaatcagc tagcggatcc accggtgtct
      841 agaaagcttt gaggccggcc tgcagggatc agcctcgact gtgccttcta gttgccagcc
      901 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
      961 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1021 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1081 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     1141 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1201 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1261 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1321 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1381 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1441 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1501 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1561 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     1621 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     1681 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1741 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     1801 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     1861 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1921 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     1981 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2041 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2101 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2161 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2221 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2281 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2341 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2401 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2461 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     2521 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2581 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2641 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2701 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2761 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     2821 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2881 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     2941 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3001 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3061 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3121 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3181 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3241 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3301 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3361 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3421 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3481 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3541 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3601 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3661 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3721 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3781 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3841 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3901 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3961 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4021 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4081 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4141 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4201 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4261 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4321 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4381 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4441 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4501 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4561 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4621 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4681 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4741 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4801 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4861 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4921 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4981 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5041 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5101 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5161 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5221 ttt
//
