LOCUS       sequence-364214-        5349 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven transmembrane domain from 
            Junctophilin-1 (TM) for membrane localization of fusion proteins 
            attached to a secretion signal Igk (PhCMV-IgkSS-TM-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5349)
  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 5349)
  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: pTS2381-Tier1-PhCMV-Igk_TM
FEATURES             Location/Qualifiers
     source          1..5349
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..38
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(155..172)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(326..914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(406..425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1085..1876,1877..1945))
                     /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     1708..1727
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1946..2050)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        2121..2500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2501..2704
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2655..2675
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2749..2768
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2749..2767
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     2808..2864
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="ETDTLLLWVLLLWVPGSTG"
     primer_bind     complement(3021..3038)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3027..3251
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3297..3725
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3384..3403)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3594..3615
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3734..3754)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3739..4068
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3919..4054
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3981..4000
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4135..4929
                     /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(4189..4208)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4799..4818
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5103..5224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5140..5159)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5194..5213
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5273..5289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5273..5289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5286..5308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5297..5313
                     /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(5321..5327,5328..5345,5346..5349,1..2))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
       61 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      121 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      181 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      241 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      301 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      361 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      421 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      481 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      541 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      601 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      661 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      721 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      781 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      841 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      901 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      961 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1021 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1081 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1141 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1201 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1261 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1321 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1381 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1441 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1501 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1561 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1621 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1681 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1741 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1801 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1861 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1921 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1981 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2041 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     2101 acgcgtggta ccctcgagtt gacattgatt attgagtagt tattaatagt aatcaattac
     2161 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     2221 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     2281 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     2341 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     2401 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     2461 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     2521 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     2581 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     2641 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     2701 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     2761 ctatagggag acccaagctg ttcgaagcgg aattcaccat gactagtgag acagacacac
     2821 tcctgctatg ggtactgctg ctctgggttc caggttccac tggtgacgct agtggtggtt
     2881 ctggttggcg ctggaacagc atcatgatcg tgctggtgat gctgctgaac atcggcctgg
     2941 ccatcctgtt cgtgcacttc ctgaccgcta gcggatccac cggtgtctag aaagctttga
     3001 ggccggcctg cagggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     3061 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     3121 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     3181 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     3241 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     3301 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3361 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3421 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3481 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3541 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3601 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3661 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3721 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3781 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3841 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3901 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3961 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4021 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4081 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4141 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4201 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4261 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4321 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4381 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4441 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4501 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4561 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4621 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4681 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4741 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4801 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4861 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4921 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     4981 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5041 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5101 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5161 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5221 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5341 taaagtgta
//
