LOCUS       sequence-451284-        7448 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses human KCNQ1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7448)
  TITLE     KCNQ1, an unexpected mediator of sex-dependent perception of 
            moderate cold temperatures
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7448)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: hKCNQ1 pCDNA3.1(-)
FEATURES             Location/Qualifiers
     source          1..7448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3039..3056)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3045..3269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3315..3743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3402..3421)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3612..3633
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3752..3772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3757..4086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3937..4072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3999..4018
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4153..4947
                     /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(4207..4226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4817..4836
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5121..5242
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5158..5177)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5212..5231
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5291..5307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5291..5307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5304..5326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5315..5331
                     /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(5339..5345,5346..5363,5364..5369))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5384..5405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5522..5539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5693..6281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5773..5792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6452..7243,7244..7312))
                     /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     7075..7094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7313..7417)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatatct gcagaattcg
      961 atctagcacc atggccgcgg cctcctcccc gcccagggcc gagaggaagc gctggggttg
     1021 gggccgcctg ccaggcgccc ggcggggcag cgcgggcctg gccaagaagt gccccttctc
     1081 gctggagctg gcggagggcg gcccggcggg cggcgcgctc tacgcgccca tcgcgcccgg
     1141 cgccccaggt cccgcgcccc ctgcgtcccc ggccgcgccc gccgcgcccc cagttgcctc
     1201 cgaccttggc ccgcggccgc cggtgagcct agacccgcgc gtctccatct acagcacgcg
     1261 ccgcccggtg ttggcgcgca cccacgtcca gggccgcgtc tacaacttcc tggagcgtcc
     1321 caccggctgg aaatgcttcg tttaccactt cgccgtcttc ctcatcgtcc tggtctgcct
     1381 catcttcagc gtgctgtcca ccatcgagca gtatgccgcc ctggccacgg ggactctctt
     1441 ctggatggag atcgtgctgg tggtgttctt cgggacggag tacgtcgtac gcctctggtc
     1501 cgccggctgc cgcagcaagt acgtgggcct ctgggggcgg ctgcgctttg cccggaagcc
     1561 catttccatc atcgatctca tcgtggtcgt ggcctccatg gtggtcctct gcgtgggctc
     1621 caaggggcag gtgtttgcca cgtcggccat caggggcatc cgcttcctgc agatcctgag
     1681 gatgctacac gtcgaccgcc agggaggcac ctggaggctc ctgggctccg tggtcttcat
     1741 ccaccgccag gagctgataa ccaccctgta catcggcttc ctgggcctca tcttctcgag
     1801 ctactttgtg tacctggctg agaaggacgc ggtgaacgag tcaggccgcg tggagttcgg
     1861 cagctacgca gatgcattgt ggtggggggt ggtcacagtc accaccatcg gctatgggga
     1921 caaggtaccc cagacgtggg tcgggaagac catcgcctcc tgcttctctg tctttgccat
     1981 ctccttcttt gcgctcccag cggggattct tggatccggg tttgccctga aggtgcagca
     2041 gaagcagagg cagaagcact tcaaccggca gatcccggcg gcagcctcac tcattcagac
     2101 cgcatggagg tgctatgctg ccgagaaccc cgactcctcc acctggaaga tctacatccg
     2161 gaaggccccc cggagccaca ctctgctgtc acccagcccc aaacccaaga agtctgtggt
     2221 ggtaaagaaa aaaaagttca agctggacaa agacaatggg gtgactcctg gagagaagat
     2281 gctcacagtc ccccatatca cgtgcgaccc cccagaagag cggcggctgg accacttctc
     2341 tgtcgacggc tatgacagtt ctgtaaggaa gagcccaaca ctgctggaag tgagcatgcc
     2401 ccatttcatg agaaccaaca gcttcgccga ggacctggac ctggaagggg agactctgct
     2461 gacacccatc acccacatct cacagctgcg ggaacaccat cgggccacca ttaaggtcat
     2521 tcgacgcatg cagtactttg tggccaagaa gaaattccag caagcgcgga agccttacga
     2581 tgtgcgggac gtcattgagc agtactcgca gggccacctc aacctcatgg tgcgcatcaa
     2641 ggagctgcag aggaggctgg accagtccat tgggaagccc tcactgttca tctccgtctc
     2701 agaaaagagc aaggatcgcg gcagcaacac gatcggcgcc cgcctgaacc gagtagaaga
     2761 caaggtgacg cagctggacc agaggctggc actcatcacc gacatgcttc accagctgct
     2821 ctccttgcac ggtggcagca cccccggcag cggcggcccc cccagagagg gcggggccca
     2881 catcacccag ccctgcggca gtggcggctc cgtcgaccct gagctattcc tgcccagcaa
     2941 caccctgccc acctacgagc agctgaccgt gcccaggagg ggccccgatg aggggtcctg
     3001 aggaggatca tcaagcttaa gtttaaaccg ctgatcagcc tcgactgtgc cttctagttg
     3061 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     3121 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     3181 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     3241 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     3301 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3361 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3421 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3481 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3541 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3601 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3661 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3721 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3781 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3841 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3901 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4021 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     4081 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     4141 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     4201 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     4261 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     4321 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     4381 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     4441 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4501 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4561 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4621 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4681 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4741 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4801 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4861 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4921 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4981 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     5041 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     5101 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     5161 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5221 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     5281 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5341 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     5401 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6121 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6361 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6421 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6481 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6541 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6601 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6661 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6721 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6781 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6841 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6901 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6961 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7021 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7081 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7141 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7201 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7261 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7321 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7381 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7441 ctgacgtc
//
