LOCUS       sequence-364993-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP expression with a 3'UTR 
            env140 ribozyme (OTetO7-PCMVmin-2-SEAP-3'UTRenv140-pA). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6563)
  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 6563)
  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: pVH267-Tier1-OTetO7-PCMVmin2-SEAP-env140
FEATURES             Location/Qualifiers
     source          1..6563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(119..135)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(119..135)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(132..154)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    143..159
                     /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(167..173,174..191,192..197))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    212..233
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(350..367)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(521..1109)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(601..620)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1280..2071,2072..2140))
                     /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     1903..1922
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2141..2245)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2327..2345
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2363..2381
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2399..2417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2435..2453
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2471..2489
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2507..2525
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2543..2561
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2615..2639
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2691..2710
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2691..2709
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4430..4447)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4436..4660
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4706..5134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4793..4812)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5003..5024
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5143..5163)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5148..5477
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5328..5463
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5390..5409
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5544..6338
                     /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(5598..5617)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6208..6227
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6512..6563,1..70)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(6549..6563,1..5))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
       61 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
      121 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      181 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      241 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      301 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
      361 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
      421 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
      481 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      541 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      601 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      661 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
      721 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
      781 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      841 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      901 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      961 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1021 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1081 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1141 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1201 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1261 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1321 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1381 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1441 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1501 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1561 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1621 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1681 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1741 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1801 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1861 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1921 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1981 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2041 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2101 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2161 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2221 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     2281 gatcgggaga tctccacgcg tccgtacacg cctaaagcat atacgttctc tatcactgat
     2341 agggagtaaa ctggatatac gttctctatc actgataggg agtaaactgt agatacgttc
     2401 tctatcactg atagggagta aactggtcat acgttctcta tcactgatag ggagtaaact
     2461 ccttatacgt tctctatcac tgatagggag taaagtctgc atacgttctc tatcactgat
     2521 agggagtaaa ctcttcatac gttctctatc actgataggg agtaaactcg agtatgtcga
     2581 ggtggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
     2641 cgagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     2701 cactataggg agacccaagc tgttcgaagc ggaattcacc atgactagtt ggctgtggct
     2761 ggtgctgctg ctgctgtgcc tggccggcaa cgtgcaggcc gctagtatca tccccgtgga
     2821 ggaggagaac cccgacttct ggaacagaga ggccgccgag gccctgggcg ccgccaagaa
     2881 gctgcagccc gcccagaccg ccgccaagaa cctgatcatc ttcctgggcg acggcatggg
     2941 cgtgagcacc gtgaccgccg ccagaatcct gaagggccag aagaaggaca agctgggccc
     3001 cgagatcccc ctggccatgg acagattccc ctacgtggcc ctgagcaaga cctacaacgt
     3061 ggacaagcac gtgcccgaca gcggcgccac cgccaccgcc tacctgtgcg gcgtgaaggg
     3121 caacttccag accatcggcc tgagcgccgc cgccagattc aaccagtgca acaccaccag
     3181 aggcaacgag gtgatcagcg tgatgaacag agccaagaag gccggcaaga gcgtgggcgt
     3241 ggtgaccacc accagagtgc agcacgccag ccccgccggc acctacgccc acaccgtgaa
     3301 cagaaactgg tacagcgacg ccgacgtgcc cgccagcgcc agacaggagg gctgccagga
     3361 catcgccacc cagctgatca gcaacatgga catcgacgtg atcctgggcg gcggcagaaa
     3421 gtacatgttc agaatgggca cccccgaccc cgagtacccc gacgactaca gccagggcgg
     3481 caccagactg gacggcaaga acctggtgca ggagtggctg gccaagagac agggcgccag
     3541 atacgtgtgg aacagaaccg agctgatgca ggccagcctg gaccccagcg tgacccacct
     3601 gatgggcctg ttcgagcccg gcgacatgaa gtacgagatc cacagagaca gcaccctgga
     3661 ccccagcctg atggagatga ccgaggccgc cctgagactg ctgagcagaa accccagagg
     3721 cttcttcctg ttcgtggagg gcggcagaat cgaccacggc caccacgaga gcagagccta
     3781 cagagccctg accgagacca tcatgttcga cgacgccatc gagagagccg gccagctgac
     3841 cagcgaggag gacaccctga gcctggtgac cgccgaccac agccacgtgt tcagcttcgg
     3901 cggctacccc ctgagaggca gcagcatctt cggcctggcc cccggcaagg ccagagacag
     3961 aaaggcctac accgtgctgc tgtacggcaa cggccccggc tacgtgctga aggacggcgc
     4021 cagacccgac gtgaccgaga gcgagagcgg cagccccgag tacagacagc agagcgccgt
     4081 gcccctggac gaggagaccc acgccggcga ggacgtggcc gtgttcgcca gaggccccca
     4141 ggcccacctg gtgcacggcg tgcaggagca gaccttcatc gcccacgtga tggccttcgc
     4201 cgcctgcctg gagccctaca ccgcctgcga cctggccccc cccgccggca ccaccgacgc
     4261 cgcccacccc ggctacagca gagtgggcgc cgccggcaga ttcgagcaga ccgctagcgg
     4321 atccaccggt gtctagaaag cttaaaccct cctcgtccgg ggctggaccg ccccgctgac
     4381 gaggcccgcg gagggccgaa acgaggaggg gccggcctgc agggatcagc ctcgactgtg
     4441 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4501 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4561 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4621 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4681 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4741 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4801 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4861 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4921 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4981 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5041 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     5101 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     5161 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5221 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5281 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5341 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5401 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5461 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5521 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5581 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5641 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5701 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5761 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5821 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5881 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5941 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     6001 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     6061 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     6121 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     6181 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     6241 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     6301 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     6361 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6421 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6481 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6541 caaataaagc aatagcatca caa
//
