LOCUS       sequence-364211-        5283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tier-1 vector encoding PhCMV-driven transmembrane domain from 
            Junctophilin-1 (TM) (PhCMV-TM-pA)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  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 5283)
  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: pTS2380-Tier1-PhCMV-TM
FEATURES             Location/Qualifiers
     source          1..5283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     716..735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        716..734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(922..939)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    928..1152
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1198..1626
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1285..1304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1495..1516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1635..1655)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1640..1969
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1820..1955
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1882..1901
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2036..2830
                     /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(2090..2109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2700..2719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3004..3125
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3041..3060)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3095..3114
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3174..3190)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3174..3190)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3187..3209)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3198..3214
                     /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(3222..3228,3229..3246,3247..3252))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3267..3288
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3405..3422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3576..4164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3656..3675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4335..5126,5127..5195))
                     /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     4958..4977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5196..5283,1..17))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
       61 gatctccacg cgtggtaccc tcgagttgac attgattatt gagtagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      721 cgactcacta tagggagacc caagctgttc gaagcggaat tcaccatgac tagtggtggt
      781 tctggttggc gctggaacag catcatgatc gtgctggtga tgctgctgaa catcggcctg
      841 gccatcctgt tcgtgcactt cctgaccgct agcggatcca ccggtgtcta gaaagctttg
      901 aggccggcct gcagggatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
      961 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1021 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1081 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1141 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1261 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1381 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1501 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1561 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1621 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1681 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1741 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1801 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1861 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     1921 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     1981 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     2041 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2101 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2161 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2221 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2281 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2341 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2401 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2461 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2521 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     2581 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2641 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2701 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2761 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2821 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     2881 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     2941 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3001 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3061 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3121 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3181 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3241 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3301 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3361 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3421 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3481 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3541 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3601 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3661 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3721 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3781 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3841 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3901 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3961 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4021 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4081 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4141 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4201 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4261 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4321 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4381 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4441 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4501 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4561 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4621 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4681 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4741 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4801 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4861 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4921 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4981 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5041 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5101 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5161 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5221 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5281 aaa
//
