LOCUS       sequence-365225-        5811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven tetracycline-activated rTetR 
            (PhCMV-rTetR-VPR-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5811)
  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 5811)
  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: pAB300-Tier1-PhCMV-rTetR
FEATURES             Location/Qualifiers
     source          1..5811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        457..660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     611..631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     705..724
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        705..723
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1439..1456)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1445..1669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1715..2143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1802..1821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2012..2033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2152..2172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2157..2486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2337..2472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2399..2418
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2553..3347
                     /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(2607..2626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3217..3236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3521..3642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3558..3577)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3612..3631
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3691..3707)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3691..3707)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3704..3726)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3715..3731
                     /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(3739..3745,3746..3763,3764..3769))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3784..3805
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3922..3939)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4093..4681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4173..4192)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4852..5643,5644..5712))
                     /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     5475..5494
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5713..5811,1..6))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctccacgc
       61 gtggtaccct cgagttgaca ttgattattg agtagttatt aatagtaatc aattacgggg
      121 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      241 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      301 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      361 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      421 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      481 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      541 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      601 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      661 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      721 agggagaccc aagctgttcg aagcggaatt caccatgact agtggatcaa gactggacaa
      781 gagcaaagtc ataaactctg ctctggaatt actcaatgga gtcggtatcg aaggcctgac
      841 gacaaggaaa ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa
      901 gaacaagcgg gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca
      961 ctcctgcccc ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcata
     1021 ccgctgtgct ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac
     1081 agagaaacag tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc
     1141 cctggagaac gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt
     1201 ggaggaacag gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat
     1261 gcccccactt ctgaaacaag caattgagct gttcgaccgg cagggagccg aacctgcctt
     1321 ccttttcggc ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg
     1381 cggggctagc ggatccaccg gtgtctagaa agctttgagg ccggcctgca gggatcagcc
     1441 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     1501 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     1561 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     1621 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     1681 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     1741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1921 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     2161 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2221 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     2281 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     2341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     2401 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     2461 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     2521 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     2581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     2701 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     2761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     2821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     2881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     2941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3061 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3361 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3421 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     3481 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     3541 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3601 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     3661 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3721 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3781 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3841 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3901 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3961 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4021 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4081 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4141 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4201 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4261 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4321 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4381 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4441 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4501 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4561 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4681 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4741 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4801 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4861 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4921 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4981 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5041 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5101 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5161 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5221 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5281 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5341 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5401 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5461 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5521 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5581 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5641 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5701 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5761 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg t
//
