LOCUS       Exported                4671 bp DNA     circular SYN 15-OCT-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          383..1690
                     /chromosome="17"
                     /map="17q24.3"
                     /mol_type="mRNA"
                     /note="color: #ffffff"
                     /db_xref="taxon:9606"
                     /organism="Homo sapiens"
     misc_feature    36..142
                     /label=ITR
                     /note="color: #a6acb3"
     primer_bind     116..135
                     /label=HA-seq-F
                     /note="color: black; sequence: tggccaactccatcactagg; added:
                     2023-02-06"
     promoter        165..376
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     375..403
                     /label=BamhI-hKCNJ2-F
                     /note="color: black; sequence: 
                     tagggatccatgggcagtgtgcgaaccaac; added: 2023-01-31"
     CDS             383..1690
                     /codon_start=1
                     /gene="KCNJ2"
                     /gene_synonym="ATFB9; HHBIRK1; HHIRK1; IRK1; KIR2.1; LQT7; 
                     SQT3"
                     /product="inward rectifier potassium channel 2"
                     /label=KCNJ2
                     /note="inward rectifier K+ channel KIR2.1; potassium 
                     inwardly-rectifying channel, subfamily J, member 2; cardiac
                     inward rectifier potassium channel; IRK-1; hIRK1; potassium
                     channel, inwardly rectifying subfamily J, member 2"
                     /note="color: #993366"
                     /db_xref="CCDS:CCDS11688.1"
                     /db_xref="GeneID:3759"
                     /db_xref="HGNC:HGNC:6263"
                     /db_xref="MIM:600681"
                     /protein_id="NP_000882.1"
                     /translation="MGSVRTNRYSIVSSEEDGMKLATMAVANGFGNGKSKVHTRQQCRS
                     RFVKKDGHCNVQFINVGEKGQRYLADIFTTCEDIRWRWMLVIFCLAFVLSWLFFGCVFW
                     LIALLHGDLDAYPYDVPDYASKEGKACVSEVNSFTAAFLFSIETQTTIGYGFRCVTDEC
                     PIAVFMVVFQSIVGCIIDAFIIGAVMAKMAKPKKRNETLVFSHNAVIAMRDGKLCLMWR
                     VGNLRKSHLVEAHVRAQLLKSRITSEGEYIPLDQIDINVGFDSGIDRIFLVSPITIVHE
                     IDEDSPLYDLSKQDIDNADFEIVVILEGMVEATAMTTQCRSSYLANEILWGHRYEPVLF
                     EEKHYYKVDYSRFHKTYEVPNTPLCSARDLAEKKYILSNANSFCYENEVALTSKEEDDS
                     ENGVPESTSTDTPPDIDLHNQASVPLEPRPLRRESEI"
     primer_bind     460..479
                     /label=B353qPCRkcnj2F1
                     /note="color: black; sequence: tgcaaatggctttgggaacg; added:
                     2023-02-06"
     primer_bind     481..501
                     /label=hKCNJ2-qPCR-F2
                     /note="color: black; sequence: gaagagtaaagtccacacccg; 
                     added: 2023-03-08"
     misc_feature    611..613
                     /label=V77E
                     /note="color: #ff0000"
     primer_bind     complement(626..645)
                     /label=hKCNJ2-qPCR-R2
                     /note="color: black; sequence: aagataaccagcatccaccg; added:
                     2023-03-08"
     primer_bind     complement(718..737)
                     /label=hKCNJ2-HA-qPCR-R
                     /note="color: black; sequence: CGTATGGGTAtgcatccagg; added:
                     2023-02-23"
     CDS             725..727
                     /label=Ala 115
                     /note="color: #ff0000; direction: RIGHT"
     CDS             728..754
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     primer_bind     complement(739..759)
                     /label=hKCNJ2-HA-qPCR-R2
                     /note="color: black; sequence: ttggaAGCGTAATCTGGAACA; 
                     added: 2023-02-28"
     primer_bind     complement(755..771)
                     /label=B353qPCRkcnj2R
                     /note="color: black; sequence: gctttgccctctttggatgc; added:
                     2023-02-06"
     primer_bind     complement(1669..1701)
                     /label=EcoRI-hKCNJ2-R
                     /note="color: black; sequence: 
                     atcgaattCTAtatctccgactctcgccgtaag; added: 2023-02-23"
     primer_bind     complement(1797..1820)
                     /label=HA-seq-R
                     /note="color: black; sequence: tgttgttaacttgtttattgcagc; 
                     added: 2023-02-06"
     misc_feature    1883..1995
                     /label=ITR delete
                     /note="color: #a6acb3"
     primer_bind     complement(2026..2042)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     promoter        2516..2620
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2621..3481
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 2621 .. 2689 / #ccffcc / signal sequence
                        2: 2690 .. 3481 / #ccffcc
                     Cleavage site after base 2689"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3652..4240
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    4528..4549
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        4564..4594
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 4564 .. 4569 / #ffffff / -35
                        2: 4570 .. 4587 / #ffffff
                        3: 4588 .. 4594 / #ffffff / -10"
     protein_bind    4602..4618
                     /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)."
                     /note="color: #31849b"
     primer_bind     4626..4642
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
ORIGIN
        1 aaagcttccc ggggggatct gggccactcc ctctctgcgc gctcgctcgc tcactgaggc
       61 cgggcgacca aaggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctggaggggt ggagtcgtga cctagggcag agcgcacatc
      181 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgatccggtg cctagagaag
      241 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
      301 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
      361 tgccgccaga acacagggat ccatgggcag tgtgcgaacc aaccgctaca gcatcgtctc
      421 ttcagaagaa gacggtatga agttggccac catggcagtt gcaaatggct ttgggaacgg
      481 gaagagtaaa gtccacaccc gacaacagtg caggagccgc tttgtgaaga aagatggcca
      541 ctgtaatgtt cagttcatca atgtgggtga gaaggggcaa cggtacctcg cagacatctt
      601 caccacgtgt gaggacattc gctggcggtg gatgctggtt atcttctgcc tggctttcgt
      661 cctgtcatgg ctgttttttg gctgtgtgtt ttggttgata gctctgctcc atggggacct
      721 ggatgcatac ccatacgatg ttccagatta cgcttccaaa gagggcaaag cttgtgtgtc
      781 cgaggtcaac agcttcacgg ctgccttcct cttctccatt gagacccaga caaccatagg
      841 ctatggtttc agatgtgtca cggatgaatg cccaattgct gttttcatgg tggtgttcca
      901 gtcaatcgtg ggctgcatca tcgatgcttt catcattggc gcagtcatgg ccaagatggc
      961 aaagccaaag aagagaaacg agactcttgt cttcagtcac aatgccgtga ttgccatgag
     1021 agacggcaag ctgtgtttga tgtggcgagt gggcaatctt cggaaaagcc acttggtgga
     1081 agctcatgtt cgagcacagc tcctcaaatc cagaattact tctgaagggg agtatatccc
     1141 tctggatcaa atagacatca atgttgggtt tgacagtgga atcgatcgta tatttctggt
     1201 gtccccaatc actatagtcc atgaaataga tgaagacagt cctttatatg atttgagtaa
     1261 acaggacatt gacaacgcag actttgaaat cgtggtcata ctggaaggca tggtggaagc
     1321 cactgccatg acgacacagt gccgtagctc ttatctagca aatgaaatcc tgtggggcca
     1381 ccgctatgag cctgtgctct ttgaagagaa gcactactac aaagtggact attccaggtt
     1441 ccacaaaact tacgaagtcc ccaacactcc cctttgtagt gccagagact tagcagaaaa
     1501 gaaatatatc ctctcaaatg caaattcatt ttgctatgaa aatgaagttg ccctcacaag
     1561 caaagaggaa gacgacagtg aaaatggagt tccagaaagc actagtacgg acacgccccc
     1621 tgacatagac cttcacaacc aggcaagtgt acctctagag cccaggccct tacggcgaga
     1681 gtcggagata tagaattcga tatcaagctt atcggcggcc gctaggcctc acctgcgatc
     1741 tcgatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1801 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1861 gtgggaggtt ttttaaacta gtccactccc tctctgcgcg ctcgctcgct cactgaggcc
     1921 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     1981 gcgcgcagag agggacagat ccgggcccgc atgcgtcgac aattcactgg ccgtcgtttt
     2041 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2101 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2161 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2221 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2281 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2341 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2401 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2461 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2521 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2581 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2641 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2701 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2761 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2821 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2881 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2941 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3001 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3061 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3121 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3181 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3241 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3301 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3361 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3421 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3481 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3541 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3601 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3661 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3721 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3781 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3841 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3901 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3961 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4021 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4081 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4141 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4201 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4261 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4321 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4381 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4441 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4501 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4561 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4621 tcacacagga aacagctatg accatgatta cgccaagctc tcgagatcta g
//
