LOCUS       sequence-364225-        5586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven blasticidin resistance gene 
            BlastR (PhCMV-BlastR-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  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 5586)
  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: pTS2402-Tier1-PhCMV-BlastR
FEATURES             Location/Qualifiers
     source          1..5586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..431
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        432..635
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     586..606
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     680..699
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        680..698
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1189..1206)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1195..1419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1465..1893
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1552..1571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1762..1783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1902..1922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1907..2236
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2087..2222
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2149..2168
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2303..3097
                     /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(2357..2376)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2967..2986
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3271..3392
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3308..3327)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3362..3381
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3441..3457)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3441..3457)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3454..3476)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3465..3481
                     /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(3489..3495,3496..3513,3514..3519))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3534..3555
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3672..3689)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3843..4431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3923..3942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4602..5393,5394..5462))
                     /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     5225..5244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5463..5567)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctgacg tcgacggatc gggagatctc cacgcgtggt accctcgagt tgacattgat
       61 tattgagtag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      121 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      181 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      241 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      301 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      361 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      421 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      481 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      541 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      601 ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg
      661 cttactggct tatcgaaatt aatacgactc actataggga gacccaagct gttcgaagcg
      721 gaattcacca tgactagtgg tggttctggt ttgtctcaag aagaatccac cctcattgaa
      781 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
      841 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
      901 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
      961 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     1021 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     1081 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggcgctagc
     1141 ggatccaccg gtgtctagaa agctttgagg ccggcctgca gggatcagcc tcgactgtgc
     1201 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1261 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1321 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1381 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     1441 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1501 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1561 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1621 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1681 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1741 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1801 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1861 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     1921 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1981 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2041 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2101 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2161 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2221 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     2281 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2341 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     2401 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     2461 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     2521 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     2581 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     2641 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     2701 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     2761 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     2821 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     2881 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     2941 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3001 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3061 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3121 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3181 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3241 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3301 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3361 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3421 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3481 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3541 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3601 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3661 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3721 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3781 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3841 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3901 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3961 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4021 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4081 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4141 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4201 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4261 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4321 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4381 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4441 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4501 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4561 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4621 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4681 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4741 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4801 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4861 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4921 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4981 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5041 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5101 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5161 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5221 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5281 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5341 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5401 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5461 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5521 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5581 aaagtg
//
