LOCUS       sequence-364204-        5305 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven ER-retention signal KDEL 
            (PhCMV-KDEL-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5305)
  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 5305)
  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: pTS2378-Tier1-PhCMV-KDEL
FEATURES             Location/Qualifiers
     source          1..5305
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..444
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        445..648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     599..619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     693..712
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        693..711
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             779..808
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(921..938)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    927..1151
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1197..1625
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1284..1303)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1494..1515
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1634..1654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1639..1968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1819..1954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1881..1900
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2035..2829
                     /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(2089..2108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2699..2718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3003..3124
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3040..3059)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3094..3113
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3173..3189)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3173..3189)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3186..3208)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3197..3213
                     /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(3221..3227,3228..3245,3246..3251))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3266..3287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3404..3421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3575..4163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3655..3674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4334..5125,5126..5194))
                     /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     4957..4976
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5195..5299)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctccacgcgt ggtaccctcg
       61 agttgacatt gattattgag tagttattaa tagtaatcaa ttacggggtc attagttcat
      121 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      181 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      241 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      301 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      361 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      421 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
      481 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
      541 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
      601 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact
      661 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
      721 gctgttcgaa gcggaattca ccatgactag tgctagcgga tccaaggacg agctgtaaga
      781 acaaaaactt atttctgaag aggatctgtg atgacgtaac aattacgggt ctaccatcga
      841 gggcctgctc gatctcccgg acgacgacgc cccgaagagg cggggtctag aaagctttga
      901 ggccggcctg cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
      961 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1021 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1081 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     1141 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     1201 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1261 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1321 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1381 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1441 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1501 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     1561 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     1621 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     1681 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     1741 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     1801 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     1861 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     1921 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     1981 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2041 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2101 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2161 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2221 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2281 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2341 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2401 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2461 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2521 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     2581 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     2641 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     2701 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     2761 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     2821 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     2881 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     2941 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3001 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3061 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3121 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     3181 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3241 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3301 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3361 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3421 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3481 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3541 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3601 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3661 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3721 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3781 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3841 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3901 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3961 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4021 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4081 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4141 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4201 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4261 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4321 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4381 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4441 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4501 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4561 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4621 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4681 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4741 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4801 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4861 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4921 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4981 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5041 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5101 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5161 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5221 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5281 aacaaatagg ggttccgcgc acatt
//
