LOCUS       sequence-364995-        6491 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP expression 
            (OTetO7-PCMVmin-1-SEAP-pA). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6491)
  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 6491)
  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: pVH255-Tier1-OTetO7-PCMVmin1-SEAP
FEATURES             Location/Qualifiers
     source          1..6491
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     47..66
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             201..995
                     /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(255..274)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     865..884
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1169..1290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1206..1225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1260..1279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1339..1355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1339..1355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1352..1374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1363..1379
                     /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(1387..1393,1394..1411,1412..1417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1432..1453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1570..1587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1741..2329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1821..1840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2500..3291,3292..3360))
                     /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     3123..3142
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3361..3465)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    3547..3565
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3583..3601
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3619..3637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3655..3673
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3691..3709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3727..3745
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3763..3781
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        3792..3829
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3826..3850
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3902..3921
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3902..3920
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5578..5595)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5584..5808
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5854..6282
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5941..5960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6151..6172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6291..6311)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6296..6491,1..134)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6476..6491,1..120)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
       61 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
      121 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
      181 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
      241 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
      301 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
      361 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
      421 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
      481 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
      541 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
      601 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
      661 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
      721 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
      781 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
      841 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
      901 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
      961 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     1021 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     1081 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     1141 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     1201 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     1261 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     1321 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1381 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1441 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1501 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1561 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     1621 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     1681 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1741 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1801 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1861 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1921 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1981 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2041 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2101 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2161 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2221 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2281 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2341 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2401 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2461 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     2521 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     2581 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     2641 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     2701 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     2761 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     2821 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     2881 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     2941 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3001 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3061 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3121 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3181 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3241 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3301 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3361 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3421 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3481 agtgccacct gacgtcgacg gatcgggaga tctccacgcg tccgtacacg cctaaagcat
     3541 atacgttctc tatcactgat agggagtaaa ctggatatac gttctctatc actgataggg
     3601 agtaaactgt agatacgttc tctatcactg atagggagta aactggtcat acgttctcta
     3661 tcactgatag ggagtaaact ccttatacgt tctctatcac tgatagggag taaagtctgc
     3721 atacgttctc tatcactgat agggagtaaa ctcttcatac gttctctatc actgataggg
     3781 agtaaactcg agtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa
     3841 ccgtcagatc gcgagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa
     3901 ttaatacgac tcactatagg gagacccaag ctgttcgaag cggaattcac catgactagt
     3961 tggctgtggc tggtgctgct gctgctgtgc ctggccggca acgtgcaggc cgctagtatc
     4021 atccccgtgg aggaggagaa ccccgacttc tggaacagag aggccgccga ggccctgggc
     4081 gccgccaaga agctgcagcc cgcccagacc gccgccaaga acctgatcat cttcctgggc
     4141 gacggcatgg gcgtgagcac cgtgaccgcc gccagaatcc tgaagggcca gaagaaggac
     4201 aagctgggcc ccgagatccc cctggccatg gacagattcc cctacgtggc cctgagcaag
     4261 acctacaacg tggacaagca cgtgcccgac agcggcgcca ccgccaccgc ctacctgtgc
     4321 ggcgtgaagg gcaacttcca gaccatcggc ctgagcgccg ccgccagatt caaccagtgc
     4381 aacaccacca gaggcaacga ggtgatcagc gtgatgaaca gagccaagaa ggccggcaag
     4441 agcgtgggcg tggtgaccac caccagagtg cagcacgcca gccccgccgg cacctacgcc
     4501 cacaccgtga acagaaactg gtacagcgac gccgacgtgc ccgccagcgc cagacaggag
     4561 ggctgccagg acatcgccac ccagctgatc agcaacatgg acatcgacgt gatcctgggc
     4621 ggcggcagaa agtacatgtt cagaatgggc acccccgacc ccgagtaccc cgacgactac
     4681 agccagggcg gcaccagact ggacggcaag aacctggtgc aggagtggct ggccaagaga
     4741 cagggcgcca gatacgtgtg gaacagaacc gagctgatgc aggccagcct ggaccccagc
     4801 gtgacccacc tgatgggcct gttcgagccc ggcgacatga agtacgagat ccacagagac
     4861 agcaccctgg accccagcct gatggagatg accgaggccg ccctgagact gctgagcaga
     4921 aaccccagag gcttcttcct gttcgtggag ggcggcagaa tcgaccacgg ccaccacgag
     4981 agcagagcct acagagccct gaccgagacc atcatgttcg acgacgccat cgagagagcc
     5041 ggccagctga ccagcgagga ggacaccctg agcctggtga ccgccgacca cagccacgtg
     5101 ttcagcttcg gcggctaccc cctgagaggc agcagcatct tcggcctggc ccccggcaag
     5161 gccagagaca gaaaggccta caccgtgctg ctgtacggca acggccccgg ctacgtgctg
     5221 aaggacggcg ccagacccga cgtgaccgag agcgagagcg gcagccccga gtacagacag
     5281 cagagcgccg tgcccctgga cgaggagacc cacgccggcg aggacgtggc cgtgttcgcc
     5341 agaggccccc aggcccacct ggtgcacggc gtgcaggagc agaccttcat cgcccacgtg
     5401 atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcccc ccccgccggc
     5461 accaccgacg ccgcccaccc cggctacagc agagtgggcg ccgccggcag attcgagcag
     5521 accgctagcg gatccaccgg tgtctagaaa gctttgaggc cggcctgcag ggatcagcct
     5581 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     5641 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5701 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     5761 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     5821 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     5881 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     5941 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6001 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6061 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6121 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6181 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     6241 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     6301 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     6361 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     6421 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     6481 gcccctaact c
//
