LOCUS       sequence-364543-        6500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PTetO7-driven SEAP expression 
            (OTetO7-PCMVmin-2-SEAP-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  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 6500)
  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: pVH227-Tier1-OTetO7-PCMVmin2-SEAP
FEATURES             Location/Qualifiers
     source          1..6500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             117..911
                     /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(171..190)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     781..800
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1085..1206
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1122..1141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1176..1195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1255..1271)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1255..1271)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1268..1290)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1279..1295
                     /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(1303..1309,1310..1327,1328..1333))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1348..1369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1486..1503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1657..2245)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1737..1756)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2416..3207,3208..3276))
                     /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     3039..3058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3277..3381)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    3463..3481
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3499..3517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3535..3553
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3571..3589
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3607..3625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3643..3661
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3679..3697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     3751..3775
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     3827..3846
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3827..3845
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5503..5520)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5509..5733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5779..6207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5866..5885)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6076..6097
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6216..6236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6221..6500,1..50)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6401..6500,1..36)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6463..6482
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
       61 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
      121 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      181 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      241 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
      301 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      361 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      421 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      481 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
      541 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
      601 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
      661 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
      721 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
      781 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
      841 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
      901 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
      961 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1021 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1081 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1141 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1201 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1261 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1321 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1381 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1441 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1501 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1561 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1621 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1681 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1741 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1801 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1861 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1921 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1981 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2041 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2101 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2161 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2221 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2281 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2341 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2401 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2461 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2521 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2581 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2641 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2701 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2761 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2821 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2881 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2941 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3001 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3061 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3121 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3181 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3241 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3301 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3361 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     3421 gggagatctc cacgcgtccg tacacgccta aagcatatac gttctctatc actgataggg
     3481 agtaaactgg atatacgttc tctatcactg atagggagta aactgtagat acgttctcta
     3541 tcactgatag ggagtaaact ggtcatacgt tctctatcac tgatagggag taaactcctt
     3601 atacgttctc tatcactgat agggagtaaa gtctgcatac gttctctatc actgataggg
     3661 agtaaactct tcatacgttc tctatcactg atagggagta aactcgagta tgtcgaggtg
     3721 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcgag
     3781 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     3841 atagggagac ccaagctgtt cgaagcggaa ttcaccatga ctagttggct gtggctggtg
     3901 ctgctgctgc tgtgcctggc cggcaacgtg caggccgcta gtatcatccc cgtggaggag
     3961 gagaaccccg acttctggaa cagagaggcc gccgaggccc tgggcgccgc caagaagctg
     4021 cagcccgccc agaccgccgc caagaacctg atcatcttcc tgggcgacgg catgggcgtg
     4081 agcaccgtga ccgccgccag aatcctgaag ggccagaaga aggacaagct gggccccgag
     4141 atccccctgg ccatggacag attcccctac gtggccctga gcaagaccta caacgtggac
     4201 aagcacgtgc ccgacagcgg cgccaccgcc accgcctacc tgtgcggcgt gaagggcaac
     4261 ttccagacca tcggcctgag cgccgccgcc agattcaacc agtgcaacac caccagaggc
     4321 aacgaggtga tcagcgtgat gaacagagcc aagaaggccg gcaagagcgt gggcgtggtg
     4381 accaccacca gagtgcagca cgccagcccc gccggcacct acgcccacac cgtgaacaga
     4441 aactggtaca gcgacgccga cgtgcccgcc agcgccagac aggagggctg ccaggacatc
     4501 gccacccagc tgatcagcaa catggacatc gacgtgatcc tgggcggcgg cagaaagtac
     4561 atgttcagaa tgggcacccc cgaccccgag taccccgacg actacagcca gggcggcacc
     4621 agactggacg gcaagaacct ggtgcaggag tggctggcca agagacaggg cgccagatac
     4681 gtgtggaaca gaaccgagct gatgcaggcc agcctggacc ccagcgtgac ccacctgatg
     4741 ggcctgttcg agcccggcga catgaagtac gagatccaca gagacagcac cctggacccc
     4801 agcctgatgg agatgaccga ggccgccctg agactgctga gcagaaaccc cagaggcttc
     4861 ttcctgttcg tggagggcgg cagaatcgac cacggccacc acgagagcag agcctacaga
     4921 gccctgaccg agaccatcat gttcgacgac gccatcgaga gagccggcca gctgaccagc
     4981 gaggaggaca ccctgagcct ggtgaccgcc gaccacagcc acgtgttcag cttcggcggc
     5041 taccccctga gaggcagcag catcttcggc ctggcccccg gcaaggccag agacagaaag
     5101 gcctacaccg tgctgctgta cggcaacggc cccggctacg tgctgaagga cggcgccaga
     5161 cccgacgtga ccgagagcga gagcggcagc cccgagtaca gacagcagag cgccgtgccc
     5221 ctggacgagg agacccacgc cggcgaggac gtggccgtgt tcgccagagg cccccaggcc
     5281 cacctggtgc acggcgtgca ggagcagacc ttcatcgccc acgtgatggc cttcgccgcc
     5341 tgcctggagc cctacaccgc ctgcgacctg gccccccccg ccggcaccac cgacgccgcc
     5401 caccccggct acagcagagt gggcgccgcc ggcagattcg agcagaccgc tagcggatcc
     5461 accggtgtct agaaagcttt gaggccggcc tgcagggatc agcctcgact gtgccttcta
     5521 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     5581 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     5641 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     5701 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     5761 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5821 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5881 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5941 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6001 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6061 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     6121 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6181 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     6241 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6301 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6361 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6421 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6481 ggccgcctct gcctctgagc
//
