LOCUS       sequence-204794-        7771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of N-terminal fused Kir7.1 in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7771)
  TITLE     Bikash Pattnaik Lab plasmids
REFERENCE   2  (bases 1 to 7771)
  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: hACTB-RFP-Kir7.1
FEATURES             Location/Qualifiers
     source          1..7771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        47..238
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(70..90)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      255..390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             439..1233
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             439..1233
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(493..512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1103..1122
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1421..1440)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1465..1512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3743..3799
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3800..4531
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(5728..5761)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     CDS             5771..5827
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    7102..7223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7139..7158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7193..7212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(7230..7685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7367..7388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7579..7598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(7712..7771,1..45)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
       61 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      601 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      961 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1381 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1441 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1501 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1561 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1621 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1681 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccatg cattagttat taataacact ggctcgtgtg acaaggccat
     2581 gaggctggtg taaagcggcc ttggagtgtg tattaagtag gcgcacagta ggtctgaaca
     2641 gactccccat cccaagaccc cagcacactt agccgtgttc tttgcacttt ctgcatgtcc
     2701 cccgtctggc ctggctgtcc ccagtggctt ccccagtgtg acatggtgca tctctgcctt
     2761 acagatcatg tttgagacct tcaacacccc agccatgtac gttgctatcc aggctgtgct
     2821 atccctgtac gcctctggcc gtaccactgg catcgtgatg gactccggtg acggggtcac
     2881 ccacactgtg cccatctacg aggggtatgc cctcccccat gccatcctgc gtctggacct
     2941 ggctggccgg gacctgactg actacctcat gaagatcctc accgagcgcg gctacagctt
     3001 caccaccacg gccgagcggg aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc
     3061 cctggacttc gagcaagaga tggccacggc tgcttccagc tcctccctgg agaagagcta
     3121 cgagctgcct gacggccagg tcatcaccat tggcaatgag cggttccgct gccctgaggc
     3181 actcttccag ccttcctccc tgggtgagtg gagactgtct cccggctctg cctgacatga
     3241 gggttacccc tcggggctgt gctgtggaag ctaagtcctg ccctcatttc cctctcaggc
     3301 atggagtcct gtggcatcca cgaaactacc ttcaactcca tcatgaagtg tgacgtggac
     3361 atccgcaaag acctgtacgc caacacagtg ctgtctggcg gcaccaccat gtaccctggc
     3421 attgccgaca ggatgcagaa ggagatcact gccctggcac ccagcacaat gaagatcaag
     3481 gtgggtgtct ttcctgcctg agctgacctg ggcaggtcag ctgtggggtc ctgtggtgtg
     3541 tggggagctg tcacatccag ggtcctcact gcctgtcccc ttccctcctc agatcattgc
     3601 tcctcctgag cgcaagtact ccgtgtggat cggcggctcc atcctggcct cgctgtccac
     3661 cttccagcag atgtggatca gcaagcagga gtatgacgag tccggcccct ccatcgtcca
     3721 ccgcaaatgc ttcggaagcg gagcgacgaa ttttagtcta ctaaaacaag cgggtgatgt
     3781 agaagaaaac cctggaccta tggtgtctaa gggcgaagag ctgattaagg agaacatgca
     3841 catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg
     3901 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg agggcggccc
     3961 tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca gaaccttcat
     4021 caaccacacc cagggcatcc ccgatttctt taagcagtcc ttccctgagg gcttcacatg
     4081 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
     4141 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg
     4201 ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc tgtaccccgc
     4261 tgacggcggc ctggaaggca gaaccgacat ggccctgaag ctcgtgggcg ggggccacct
     4321 gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc
     4381 cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca aagagaccta
     4441 cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca
     4501 caaacttaat ggcatggacg agctgtacaa gggcaaaaag aaaggccgga ctcaaaagga
     4561 gaagaaagct gctcgtgctc gttccaaagg gaaaaactcg gatgaagaag cccccaagac
     4621 tgcccagaag aagttaaaga tggacagcag taattgcaaa gttattgctc ctctcctaag
     4681 tcaaagatac cggaggatgg tcaccaagga tggccacagc acacttcaaa tggatggcgc
     4741 tcaaagaggt cttgcatatc ttcgagatgc ttggggaatc ctaatggaca tgcgctggcg
     4801 ttggatgatg ttggtctttt ctgcttcttt tgttgtccac tggcttgtct ttgcagtgct
     4861 ctggtatgtt ctggctgaga tgaatggtga tctggaacta gatcatgatg ccccacctga
     4921 aaaccacact atctgtgtca agtatatcac cagtttcaca gctgcattct ccttctccct
     4981 ggagacacaa ctcacaattg gttatggtac catgttcccc agtggtgact gtccaagtgc
     5041 aatcgcctta cttgccatac aaatgctcct aggcctcatg ctagaggctt ttatcacagg
     5101 tgcttttgtg gcgaagattg cccggccaaa aaatcgagct ttttcaattc gctttactga
     5161 cacagcagta gtagctcaca tggatggcaa acctaatctt atcttccaag tggccaacac
     5221 ccgacctagc cctctaacca gtgtccgggt ctcagctgta ctctatcagg aaagagaaaa
     5281 tggcaaactc taccagacca gtgtggattt ccaccttgat ggcatcagtt ctgacgaatg
     5341 tccattcttc atctttccac taacgtacta tcactccatt acaccatcaa gtcctctggc
     5401 tactctgctc cagcatgaaa atccttctca ctttgaatta gttgtattcc tttcagcaat
     5461 gcaggagggc actggagaaa tatgccaaag gaggacatcc tacctaccgt ctgaaatcat
     5521 gttacatcac tgttttgcat ctctgttgac ccgaggttcc aaaggtgaat atcaaatcaa
     5581 gatggagaat tttgacaaga ctgtccctga atttccaact cctctggttt ctaaaagccc
     5641 aaacaggact gacctggata tccacatcaa tggacaaagc attgacaatt ttcagatctc
     5701 tgaaacagga ctgacagaat aactcttata acttcgtata ggatacttta tacgaagtta
     5761 tggaagcgga gcgacgaatt ttagtctact gaaacaagcg ggagacgtgg aggaaaaccc
     5821 tggacctacc ggttgcggcc gcctcgagtc tagaactagt tgatcatagg cggactatga
     5881 cttagttgcg ttacaccctt tcttgacaaa acctaacttg cgcagaaaac aagatgagat
     5941 tggcatggct ttatttgttt tttttgtttt gttttggttt tttttttttt tttggcttga
     6001 ctcaggattt aaaaactgga acggtgaagg tgacagcagt cggttggagc gagcatcccc
     6061 caaagttcac aatgtggccg aggactttga ttgcacattg ttgttttttt aatagtcatt
     6121 ccaaatatga gatgcattgt tacaggaagt cccttgccat cctaaaagcc accccacttc
     6181 tctctaagga gaatggccca gtcctctccc aagtccacac aggggaggtg atagcattgc
     6241 tttcgtgtaa attatgtaat gcaaaatttt tttaatcttc gccttaatac ttttttattt
     6301 tgttttattt tgaatgatga gccttcgtgc ccccccttcc cccttttttg tcccccaact
     6361 tgagatgtat gaaggctttt ggtctccctg ggagtgggtg gaggcagcca gggcttacct
     6421 gtacactgac ttgagaccag ttgaataaaa gtgcacacct taaaaatgag gccaagtgtg
     6481 actttgtggt gtggctgggt tgggggcagc agagggtgaa ccctgcagga gggtgaaccc
     6541 tgcaaaaggg tggggcagtg ggggccaact tgtcctcacc cagagtgcag gtgtgtggag
     6601 atccctcctg ccttgacatt gagcagcctt agagggtggg ggaggctcag gggtcaggtc
     6661 tctgttcctg cttattgggg agttcctggc ctggcccttc tatgtctccc caggtacccc
     6721 agtttttctg ggttcaccca gagtgcagat gcttgaggag gtgggaaggg actatttggg
     6781 ggtgtctggc tcaggtgcca tgcctcactg gggctggttg gcacctgcat ttcctgggag
     6841 tggggctgtc tcagggtagc tgggcacggt gttcccttga gtgggggtgt agtgagtgtt
     6901 cctagctgcc acgcctttgc cttcacctat gggatcgtgg ctgtcggcct tgagggtcag
     6961 cctggcccag gctcccatag gcttaggaga ggccgcaatt cctacctgtt catccagaca
     7021 gagggggacc tggaatcaaa gtcaagttgg ggtaggggcc ccctgaacct gaaacataaa
     7081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     7141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7201 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     7261 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7321 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7381 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7441 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7501 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7561 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     7621 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7681 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     7741 aatacattca aatatgtatc cgctcatgag a
//
