LOCUS       sequence-217317-        8097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Kv2.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8097)
  TITLE     Sesti Kv2.1 plasmid
REFERENCE   2  (bases 1 to 8097)
  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"
COMMENT     Sequence Label: hKv2.1-V5
FEATURES             Location/Qualifiers
     source          1..8097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     163..183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          373..505
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     488..507
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        550..568
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3284..3405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3295..3314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3349..3368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3591..4046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3888..3909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4163..4520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4163..4354
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4186..4206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4371..4506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4571..5365
                     /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(4625..4644)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5235..5254
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5429..5477
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5518..5537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5637..5659
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5697..5715)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5783..5887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5888..5956,5957..6748)
                     /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     complement(6106..6125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6919..7507
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7408..7427
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7718..8097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctgcgatcgc ccgccccgtt gacgcaaatg ggcggtaggc
      181 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcactagaa
      241 gctttattgc ggtagtttat cacagttaaa ttgctaacgc agtcagtgct tctgacacaa
      301 cagtctcgaa cttaagctgc agtgactctc ttaaggtagc cttgcagaag ttggtcgtga
      361 ggcactgggc aggtaagtat caaggttaca agacaggttt aaggagacca atagaaactg
      421 ggcttgtcga gacagagaag actcttgcgt ttctgatagg cacctattgg tcttactgac
      481 atccactttg cctttctctc cacaggtgtc cactcccagt tcaattacag ctcttaaggc
      541 tagagtactt aatacgactc actataggct agcctcgaga attccaccat gccggcgggc
      601 atgacgaagc atggctcccg ctccaccagc tcgctgccgc ccgagcccat ggagatcgtg
      661 cgcagcaagg cgtgctctcg gcgggtccgc ctcaacgtcg gggggctggc gcacgaggta
      721 ctctggcgta ccctggaccg cctgccccgc acgcggctgg gcaagctccg cgactgcaac
      781 acgcacgact cgctgctcga ggtgtgcgat gactacagcc tcgacgacaa cgagtacttc
      841 tttgaccgcc acccgggcgc cttcacctcc atcctcaact tctaccgcac tgggcgactg
      901 cacatgatgg aggagatgtg cgcgctcagc ttcagccaag agctcgacta ctggggcatc
      961 gacgagatct acccggagtc ctgctgccag gcccgctacc accagaagaa agagcagatg
     1021 aacgaggagc tcaagcgtga ggccgagacc ctacgggagc gggaaggcga ggagttcgat
     1081 aacacgtgct gcgcagagaa gaggaaaaaa ctctgggacc tactggagaa gcccaattcc
     1141 tctgtggctg ccaagatcct tgccataatt tccatcatgt tcatcgtcct ctccaccatt
     1201 gccctgtccc tcaacacgct gcctgagcta cagagcctcg atgagttcgg ccagtccaca
     1261 gacaaccccc agctggccca cgtggaggcc gtgtgcatcg catggttcac catggagtac
     1321 ctgctgaggt tcctctcctc gcccaagaag tggaagttct tcaagggccc actcaatgcc
     1381 attgacttgt tggccattct gccatactat gtcaccattt tcctcaccga atccaacaag
     1441 agcgtgctgc aattccagaa tgtccgccgc gtggtccaga tcttccgcat catgcgcatt
     1501 ctccgcatcc ttaagcttgc acgccactcc actggcctcc agtctctggg cttcactttg
     1561 cggaggagct acaatgagtt gggcttgctc atcctcttcc ttgccatggg cattatgatc
     1621 ttctccagcc ttgtcttctt tgctgagaag gatgaggacg acaccaagtt caaaagcatc
     1681 ccagcctctt tctggtgggc caccatcacc atgactactg ttgggtatgg agacatctac
     1741 cccaagactc tcctggggaa aattgttggg ggactctgct gcattgcagg agtcctggtg
     1801 attgctcttc ccatccccat catcgtcaat aacttctctg agttctataa ggagcagaag
     1861 agacaggaga aagcaatcaa acggcgagag gctctggaga gagccaagag gaatggcagc
     1921 atcgtatcca tgaacatgaa ggatgctttt gcccggagca ttgagatgat ggacattgtg
     1981 gttgagaaaa atggggagaa tatgggtaag aaagacaaag tacaagataa ccacttgtct
     2041 cctaacaaat ggaaatggac aaagaggaca ctgtctgaaa ccagctcaag taagtccttt
     2101 gaaaccaagg aacagggatc ccctgaaaaa gccagatcgt cttctagtcc tcagcacctg
     2161 aacgttcagc agttggaaga catgtacaat aagatggcca agacccaatc ccaacccatc
     2221 ctcaatacca aggagtcagc agcacagagc aaaccaaagg aagaacttga aatggagagt
     2281 atccccagcc ccgtagcccc tctgcccact cgcacagaag gggtcattga catgcgaagt
     2341 atgtcaagca ttgatagttt cattagctgt gccacagact tccctgaggc caccagattc
     2401 tcccacagcc ctttgacatc actccccagc aagactgggg gcagcacagc cccagaagtg
     2461 ggctggcggg gagctctggg tgccagtggt ggtaggtttg tggaggccaa ccccagccct
     2521 gatgccagcc agcactctag tttcttcatc gagagcccca agagttccat gaaaactaac
     2581 aaccctttga agctccgagc acttaaagtc aacttcatgg agggtgaccc cagtccactc
     2641 ctccccgttc tagggatgta ccatgaccct ctcaggaacc gggggagtgc tgcggctgct
     2701 gtcgctggac tggagtgtgc cacgcttttg gacaaggctg tgctgagccc agagtcctcc
     2761 atctacacca cagcaagtgc taagacaccc ccccggtctc ctgagaaaca cacagcaata
     2821 gcgttcaact ttgaggcggg tgtccaccag tacattgacg cagacacaga tgatgaggga
     2881 cagctgctct acagtgtgga ctccagcccc cccaaaagcc tccctgggag caccagtccg
     2941 aagttcagca cggggacaag atcggagaaa aaccactttg aaagctcccc tttacccacc
     3001 tcccctaagt tcttaaggca gaactgtatt tactccacag aagcattgac tggaaaaggc
     3061 cccagtggtc aggaaaagtg caaacttgag aaccacatct cccctgacgt ccgtgtgttg
     3121 ccagggggag gagcccatgg aagcacacga gatcagagca tcgaattcac gcgtggtacc
     3181 tctagagtcg acggcaaacc aatcccaaac ccactgctgg gcctggatta aggatccgcg
     3241 gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata cattgatgag
     3301 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     3361 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     3421 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     3481 ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat agcgaagagg
     3541 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     3601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3661 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3781 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3901 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3961 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4021 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     4081 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     4141 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4201 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4261 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4381 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4441 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4501 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     4561 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4621 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4681 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4741 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4801 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4861 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4921 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4981 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5041 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5101 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5161 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5221 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5281 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5341 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5401 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5461 tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca ctctcagtac
     5521 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     5581 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     5641 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     5701 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     5761 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5821 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5881 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5941 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6001 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6061 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7561 tcacatggct cgacagatct tcaatattgg ccattagcca tattattcat tggttatata
     7621 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     7681 atttatattg gctcatgtcc aatatgaccg ccatgttggc attgattatt gactagttat
     7741 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     7801 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     7861 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     7921 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg
     7981 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     8041 ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
