LOCUS       sequence-366602-        6453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP (OTetO7-PCMVmin-SEAP-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6453)
  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 6453)
  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: pTS1017-Tier1-OTetO7-PCMVmin-SEAP
FEATURES             Location/Qualifiers
     source          1..6453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1050..1067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1056..1280
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1326..1754
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1413..1432)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1623..1644
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1763..1783)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1768..2097
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1948..2083
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2010..2029
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2164..2958
                     /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(2218..2237)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2828..2847
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3132..3253
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3169..3188)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3223..3242
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3302..3318)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3302..3318)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3315..3337)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3326..3342
                     /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(3350..3356,3357..3374,3375..3380))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3395..3416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3533..3550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3704..4292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3784..3803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4463..5254,5255..5323))
                     /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     5086..5105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5324..5428)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    5510..5528
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5546..5564
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5582..5600
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5618..5636
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5654..5672
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5690..5708
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5726..5744
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        5756..5794
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5839..5858
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5839..5857
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acgccgacgt gcccgccagc gccagacagg agggctgcca ggacatcgcc acccagctga
       61 tcagcaacat ggacatcgac gtgatcctgg gcggcggcag aaagtacatg ttcagaatgg
      121 gcacccccga ccccgagtac cccgacgact acagccaggg cggcaccaga ctggacggca
      181 agaacctggt gcaggagtgg ctggccaaga gacagggcgc cagatacgtg tggaacagaa
      241 ccgagctgat gcaggccagc ctggacccca gcgtgaccca cctgatgggc ctgttcgagc
      301 ccggcgacat gaagtacgag atccacagag acagcaccct ggaccccagc ctgatggaga
      361 tgaccgaggc cgccctgaga ctgctgagca gaaaccccag aggcttcttc ctgttcgtgg
      421 agggcggcag aatcgaccac ggccaccacg agagcagagc ctacagagcc ctgaccgaga
      481 ccatcatgtt cgacgacgcc atcgagagag ccggccagct gaccagcgag gaggacaccc
      541 tgagcctggt gaccgccgac cacagccacg tgttcagctt cggcggctac cccctgagag
      601 gcagcagcat cttcggcctg gcccccggca aggccagaga cagaaaggcc tacaccgtgc
      661 tgctgtacgg caacggcccc ggctacgtgc tgaaggacgg cgccagaccc gacgtgaccg
      721 agagcgagag cggcagcccc gagtacagac agcagagcgc cgtgcccctg gacgaggaga
      781 cccacgccgg cgaggacgtg gccgtgttcg ccagaggccc ccaggcccac ctggtgcacg
      841 gcgtgcagga gcagaccttc atcgcccacg tgatggcctt cgccgcctgc ctggagccct
      901 acaccgcctg cgacctggcc ccccccgccg gcaccaccga cgccgcccac cccggctaca
      961 gcagagtggg cgccgccggc agattcgagc agaccgctag cggatccacc ggtgtctaga
     1021 aagctttgag gccggcctgc agggatcagc ctcgactgtg ccttctagtt gccagccatc
     1081 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1141 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1201 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     1261 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     1321 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1381 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1441 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1501 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1561 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1621 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1681 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1741 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     1801 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     1861 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1921 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     1981 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2041 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     2101 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     2161 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     2221 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     2281 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     2341 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     2401 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     2461 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     2521 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     2581 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     2641 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     2701 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     2761 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     2821 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     2881 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     2941 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     3001 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     3061 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     3121 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3181 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3241 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     3301 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3361 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3421 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3481 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3541 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3601 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3661 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3721 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3781 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3841 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3901 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3961 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4021 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4081 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4141 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4201 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4261 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4321 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4381 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4441 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4501 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4561 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4621 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4681 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4741 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4801 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4861 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4921 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4981 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5041 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5101 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5161 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5221 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5281 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5341 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5401 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     5461 acggatcggg agatctccac gcgtccgtac acgcctaaag catatacgtt ctctatcact
     5521 gatagggagt aaactggata tacgttctct atcactgata gggagtaaac tgtagatacg
     5581 ttctctatca ctgataggga gtaaactggt catacgttct ctatcactga tagggagtaa
     5641 actccttata cgttctctat cactgatagg gagtaaagtc tgcatacgtt ctctatcact
     5701 gatagggagt aaactcttca tacgttctct atcactgata gggagtaaac tcgagggtag
     5761 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     5821 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg ttcgaagcgg
     5881 aattcaccat gactagtctg ctgctgctgc tgctgggcct gagactgcag ctgagcctgg
     5941 gcatcatccc cgtggaggag gagaaccccg acttctggaa cagagaggcc gccgaggccc
     6001 tgggcgccgc caagaagctg cagcccgccc agaccgccgc caagaacctg atcatcttcc
     6061 tgggcgacgg catgggcgtg agcaccgtga ccgccgccag aatcctgaag ggccagaaga
     6121 aggacaagct gggccccgag atccccctgg ccatggacag attcccctac gtggccctga
     6181 gcaagaccta caacgtggac aagcacgtgc ccgacagcgg cgccaccgcc accgcctacc
     6241 tgtgcggcgt gaagggcaac ttccagacca tcggcctgag cgccgccgcc agattcaacc
     6301 agtgcaacac caccagaggc aacgaggtga tcagcgtgat gaacagagcc aagaaggccg
     6361 gcaagagcgt gggcgtggtg accaccacca gagtgcagca cgccagcccc gccggcacct
     6421 acgcccacac cgtgaacaga aactggtaca gcg
//
