LOCUS       sequence-364235-        5556 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven N-terminal fragment of a split 
            tabacco etch virus protease splitTEV (PhCMV-N-TEV-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5556)
  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 5556)
  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: pTS2406-Tier1-PhCMV-NTEV
FEATURES             Location/Qualifiers
     source          1..5556
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     741..760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        741..759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1220..1237)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1226..1450
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1496..1924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1583..1602)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1793..1814
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1933..1953)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1938..2267
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2118..2253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2180..2199
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2334..3128
                     /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(2388..2407)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2998..3017
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3302..3423
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3339..3358)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3393..3412
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3472..3488)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3472..3488)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3485..3507)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3496..3512
                     /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(3520..3526,3527..3544,3545..3550))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3565..3586
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3703..3720)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3874..4462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3954..3973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4633..5424,5425..5493))
                     /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     5256..5275
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5494..5556,1..42))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
       61 gccacctgac gtcgacggat cgggagatct ccacgcgtgg taccctcgag ttgacattga
      121 ttattgagta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
      721 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tgttcgaagc
      781 ggaattcacc atgactagtg gtggttctgg tggagaatgc ttgtttaagg ggccgcgtga
      841 ttacaacccg atatcgagca ccatttgtca tttgacgaat gaatctgatg ggcacacaac
      901 atcgttgtat ggtattggat ttggtccctt catcattaca aacaagcact tgtttagaag
      961 aaataatgga acactgttgg tccaatcact acatggtgta ttcaaggtca agaacaccac
     1021 gactttgcaa caacacctca ttgatgggag ggacatgata attattcgca tgcctaagga
     1081 tttcccacca tttcctcaaa agctgaaatt tagagagcca caaagggaag agcgcatatg
     1141 tcttgtgaca accaacttcc aaactgctag cggatccacc ggtgtctaga aagctttgag
     1201 gccggcctgc agggatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     1261 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     1321 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     1381 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     1441 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     1501 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     1561 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     1621 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     1681 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     1741 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     1801 ttgttccaaa ctggaacaac actcmaccct atctcggtct attcttttga tttataaggg
     1861 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     1921 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     1981 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     2041 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     2101 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2161 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     2221 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     2281 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     2341 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     2401 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     2461 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     2521 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     2581 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     2641 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     2701 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     2761 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     2821 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     2881 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     2941 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3001 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3061 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3121 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3181 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3241 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3301 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     3361 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     3421 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     3481 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3541 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3601 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3661 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4501 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4561 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4621 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4681 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4741 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4801 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4861 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4921 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4981 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5041 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5101 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5161 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5221 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5281 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5341 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5401 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5461 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5521 catttatcag ggttattgtc tcatgagcgg atacat
//
