LOCUS       sequence-364997-        5316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven F-type transactivator domain 
            (PhCMV-F-type-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5316)
  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 5316)
  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: pTS2366-Tier1-PhCMV_F-type
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        446..649
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     600..620
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     694..713
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        694..712
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             762..872
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     primer_bind     complement(933..950)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    939..1163
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1209..1637
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1296..1315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1506..1527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1646..1666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1651..1980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1831..1966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1893..1912
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2047..2841
                     /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(2101..2120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2711..2730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3015..3136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3106..3125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3185..3201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3185..3201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3198..3220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3209..3225
                     /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(3233..3239,3240..3257,3258..3263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3278..3299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3416..3433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3587..4175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3667..3686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4346..5137,5138..5206))
                     /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     4969..4988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5207..5311)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctccacgcg tggtaccctc
       61 gagttgacat tgattattga gtagttatta atagtaatca attacggggt cattagttca
      121 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      181 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      301 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      361 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      421 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      481 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      541 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      601 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac
      661 tagagaaccc actgcttact ggcttatcga aattaatacg actcactata gggagaccca
      721 agctgttcga agcggaattc accatgacta gtgcaccggc cgacgccctt gacgattttg
      781 acttagacat gctcccagcc gatgcccttg acgactttga ccttgatatg ctgcctgctg
      841 acgctcttga cgattttgac cttgacatgc tccccggggc tagcggatcc accggtgtct
      901 agaaagcttt gaggccggcc tgcagggatc agcctcgact gtgccttcta gttgccagcc
      961 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1021 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1081 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1141 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     1201 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1261 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1321 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1381 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1441 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1501 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     1561 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1621 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     1681 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     1741 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1801 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     1861 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     1921 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     1981 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     2041 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2101 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     2161 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     2221 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     2281 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     2341 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     2401 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     2461 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     2521 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     2581 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     2641 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     2701 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     2761 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     2821 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     2881 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2941 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3001 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3061 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3121 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3181 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3241 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3301 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3361 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4021 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5281 tttagaaaaa taaacaaata ggggttccgc gcacat
//
