LOCUS       sequence-365013-        6463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP expression 
            (OTetO7-Pmin-SEAP-pA). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6463)
  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 6463)
  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: pVH254-Tier1-OTetO7-Pmin-SEAP
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..25
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(85..101)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(85..101)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(98..120)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    109..125
                     /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(133..139,140..157,158..163))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    178..199
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(316..333)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(487..1075)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(567..586)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1246..2037,2038..2106))
                     /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     1869..1888
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2107..2211)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    2293..2311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2329..2347
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2365..2383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2401..2419
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2437..2455
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2473..2491
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2509..2527
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        2539..2570
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     primer_bind     2620..2639
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2620..2638
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4296..4313)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4302..4526
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4572..5000
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4659..4678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4869..4890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5009..5029)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5014..5343
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5194..5329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5256..5275
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5410..6204
                     /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(5464..5483)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6074..6093
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    join(6378..6463,1..36)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6415..6434)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
       61 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
      121 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
      181 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
      241 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      301 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      361 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      901 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1201 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1261 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1321 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1381 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1441 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1501 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1561 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1621 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1681 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1741 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1801 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1861 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1921 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1981 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2041 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2101 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2161 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2221 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc cacgcgtccg tacacgccta
     2281 aagcatatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
     2341 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
     2401 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
     2461 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
     2521 atagggagta aactcgagta gagggtatat aatggaagct cgacttccag gagctctctg
     2581 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2641 gacccaagct gttcgaagcg gaattcacca tgactagttg gctgtggctg gtgctgctgc
     2701 tgctgtgcct ggccggcaac gtgcaggccg ctagtatcat ccccgtggag gaggagaacc
     2761 ccgacttctg gaacagagag gccgccgagg ccctgggcgc cgccaagaag ctgcagcccg
     2821 cccagaccgc cgccaagaac ctgatcatct tcctgggcga cggcatgggc gtgagcaccg
     2881 tgaccgccgc cagaatcctg aagggccaga agaaggacaa gctgggcccc gagatccccc
     2941 tggccatgga cagattcccc tacgtggccc tgagcaagac ctacaacgtg gacaagcacg
     3001 tgcccgacag cggcgccacc gccaccgcct acctgtgcgg cgtgaagggc aacttccaga
     3061 ccatcggcct gagcgccgcc gccagattca accagtgcaa caccaccaga ggcaacgagg
     3121 tgatcagcgt gatgaacaga gccaagaagg ccggcaagag cgtgggcgtg gtgaccacca
     3181 ccagagtgca gcacgccagc cccgccggca cctacgccca caccgtgaac agaaactggt
     3241 acagcgacgc cgacgtgccc gccagcgcca gacaggaggg ctgccaggac atcgccaccc
     3301 agctgatcag caacatggac atcgacgtga tcctgggcgg cggcagaaag tacatgttca
     3361 gaatgggcac ccccgacccc gagtaccccg acgactacag ccagggcggc accagactgg
     3421 acggcaagaa cctggtgcag gagtggctgg ccaagagaca gggcgccaga tacgtgtgga
     3481 acagaaccga gctgatgcag gccagcctgg accccagcgt gacccacctg atgggcctgt
     3541 tcgagcccgg cgacatgaag tacgagatcc acagagacag caccctggac cccagcctga
     3601 tggagatgac cgaggccgcc ctgagactgc tgagcagaaa ccccagaggc ttcttcctgt
     3661 tcgtggaggg cggcagaatc gaccacggcc accacgagag cagagcctac agagccctga
     3721 ccgagaccat catgttcgac gacgccatcg agagagccgg ccagctgacc agcgaggagg
     3781 acaccctgag cctggtgacc gccgaccaca gccacgtgtt cagcttcggc ggctaccccc
     3841 tgagaggcag cagcatcttc ggcctggccc ccggcaaggc cagagacaga aaggcctaca
     3901 ccgtgctgct gtacggcaac ggccccggct acgtgctgaa ggacggcgcc agacccgacg
     3961 tgaccgagag cgagagcggc agccccgagt acagacagca gagcgccgtg cccctggacg
     4021 aggagaccca cgccggcgag gacgtggccg tgttcgccag aggcccccag gcccacctgg
     4081 tgcacggcgt gcaggagcag accttcatcg cccacgtgat ggccttcgcc gcctgcctgg
     4141 agccctacac cgcctgcgac ctggcccccc ccgccggcac caccgacgcc gcccaccccg
     4201 gctacagcag agtgggcgcc gccggcagat tcgagcagac cgctagcgga tccaccggtg
     4261 tctagaaagc tttgaggccg gcctgcaggg atcagcctcg actgtgcctt ctagttgcca
     4321 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4381 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     4441 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     4501 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     4561 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4921 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4981 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     5041 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     5101 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5161 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5221 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5281 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5341 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     5401 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5461 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5521 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5581 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5641 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5701 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5761 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5821 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5881 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5941 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6001 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6061 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6121 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6181 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6241 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6301 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     6361 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     6421 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttc
//
