LOCUS       sequence-365011-        6534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP expression with a 
            suppressor 5'UTR (OTetO7-PCMVmin-2-5'UTRTS-SEAP-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6534)
  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 6534)
  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: pVH265-Tier1-OTetO7-PCMVmin2-TS-SEAP
FEATURES             Location/Qualifiers
     source          1..6534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     66..85
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(145..161)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(145..161)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(158..180)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    169..185
                     /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(193..199,200..217,218..223))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    238..259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(376..393)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(547..1135)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(627..646)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1306..2097,2098..2166))
                     /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     1929..1948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2167..2271)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2353..2371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2389..2407
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2425..2443
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2461..2479
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2497..2515
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2533..2551
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2569..2587
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2641..2665
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     2717..2736
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2717..2735
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4427..4444)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4433..4657
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4703..5131
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4790..4809)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5000..5021
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5140..5160)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5145..5474
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5325..5460
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5387..5406
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5541..6335
                     /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(5595..5614)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6205..6224
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6509..6534,1..96)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
       61 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
      121 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      181 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      241 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      301 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      361 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      421 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      481 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      541 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      601 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      661 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      721 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      781 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      841 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      901 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      961 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1021 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1081 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1141 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1201 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1261 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1321 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1381 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1441 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1501 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1561 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1621 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1681 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1741 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1801 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1861 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1921 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1981 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2041 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2101 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2161 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2221 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2281 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc cacgcgtccg tacacgccta
     2341 aagcatatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
     2401 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
     2461 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
     2521 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
     2581 atagggagta aactcgagta tgtcgaggtg gcgtgtacgg tgggaggcct atataagcag
     2641 agctcgttta gtgaaccgtc agatcgcgag ctctctggct aactagagaa cccactgctt
     2701 actggcttat cgaaattaat acgactcact atagggagac ccaagctgtt cgaagcggaa
     2761 ttcgagctcg cccggggaac ctctagagtc agcttcctgc atgactagtt ggctgtggct
     2821 ggtgctgctg ctgctgtgcc tggccggcaa cgtgcaggcc gctagtatca tccccgtgga
     2881 ggaggagaac cccgacttct ggaacagaga ggccgccgag gccctgggcg ccgccaagaa
     2941 gctgcagccc gcccagaccg ccgccaagaa cctgatcatc ttcctgggcg acggcatggg
     3001 cgtgagcacc gtgaccgccg ccagaatcct gaagggccag aagaaggaca agctgggccc
     3061 cgagatcccc ctggccatgg acagattccc ctacgtggcc ctgagcaaga cctacaacgt
     3121 ggacaagcac gtgcccgaca gcggcgccac cgccaccgcc tacctgtgcg gcgtgaaggg
     3181 caacttccag accatcggcc tgagcgccgc cgccagattc aaccagtgca acaccaccag
     3241 aggcaacgag gtgatcagcg tgatgaacag agccaagaag gccggcaaga gcgtgggcgt
     3301 ggtgaccacc accagagtgc agcacgccag ccccgccggc acctacgccc acaccgtgaa
     3361 cagaaactgg tacagcgacg ccgacgtgcc cgccagcgcc agacaggagg gctgccagga
     3421 catcgccacc cagctgatca gcaacatgga catcgacgtg atcctgggcg gcggcagaaa
     3481 gtacatgttc agaatgggca cccccgaccc cgagtacccc gacgactaca gccagggcgg
     3541 caccagactg gacggcaaga acctggtgca ggagtggctg gccaagagac agggcgccag
     3601 atacgtgtgg aacagaaccg agctgatgca ggccagcctg gaccccagcg tgacccacct
     3661 gatgggcctg ttcgagcccg gcgacatgaa gtacgagatc cacagagaca gcaccctgga
     3721 ccccagcctg atggagatga ccgaggccgc cctgagactg ctgagcagaa accccagagg
     3781 cttcttcctg ttcgtggagg gcggcagaat cgaccacggc caccacgaga gcagagccta
     3841 cagagccctg accgagacca tcatgttcga cgacgccatc gagagagccg gccagctgac
     3901 cagcgaggag gacaccctga gcctggtgac cgccgaccac agccacgtgt tcagcttcgg
     3961 cggctacccc ctgagaggca gcagcatctt cggcctggcc cccggcaagg ccagagacag
     4021 aaaggcctac accgtgctgc tgtacggcaa cggccccggc tacgtgctga aggacggcgc
     4081 cagacccgac gtgaccgaga gcgagagcgg cagccccgag tacagacagc agagcgccgt
     4141 gcccctggac gaggagaccc acgccggcga ggacgtggcc gtgttcgcca gaggccccca
     4201 ggcccacctg gtgcacggcg tgcaggagca gaccttcatc gcccacgtga tggccttcgc
     4261 cgcctgcctg gagccctaca ccgcctgcga cctggccccc cccgccggca ccaccgacgc
     4321 cgcccacccc ggctacagca gagtgggcgc cgccggcaga ttcgagcaga ccgctagcgg
     4381 atccaccggt gtctagaaag ctttgaggcc ggcctgcagg gatcagcctc gactgtgcct
     4441 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     4501 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     4561 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     4621 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     4681 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4741 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     4801 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4861 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     4921 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4981 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5041 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5101 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     5161 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5221 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5281 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5341 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5401 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5461 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     5521 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5581 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5641 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5701 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5761 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5821 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5881 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5941 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     6001 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     6061 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     6121 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     6181 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     6241 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     6301 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     6361 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     6421 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6481 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atgg
//
