LOCUS       Exported                7299 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses human KCNB1 in mammalian cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    hKv2.1-pEGFP-C1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  TITLE     KCNB1 unpublished plasmids
REFERENCE   2  (bases 1 to 7299)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7299
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(236..340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      367..822
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(454..473)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     664..685
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    829..950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(840..859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     894..913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3708..4424)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(3708..4424)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3750..3771)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4099..4118
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4358..4379
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4421..4430
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(4469..4672)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4498..4518)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4673..4976
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5151..5739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5231..5250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6068..6115
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6140..6159
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6347..7141)
                     /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(6458..6477)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7068..7087
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(7176..234)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7190..26
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7244..7263)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
      241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
      301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
      361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcgtta agatacattg
      841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
      901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
      961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1021 aaaacctcta caaatgtggt atggctgatt atgatcagtt atctagatcc ggtggatccc
     1081 gggcccgcgg taccgtgatg ctctgatctc gtgtgcttcc atgggctcct ccccctggca
     1141 acacacggac gtcaggggag atgtggttct caagtttgca cttttcctga ccactggggc
     1201 cttttccagt caatgcttct gtggagtaaa tacagttctg ccttaagaac ttaggggagg
     1261 tgggtaaagg ggagctttca aagtggtttt tctccgatct tgtccccgtg ctgaacttcg
     1321 gactggtgct cccagggagg cttttggggg ggctggagtc cacactgtag agcagctgtc
     1381 cctcatcatc tgtgtctgcg tcaatgtact ggtggacacc cgcctcaaag ttgaacgcta
     1441 ttgctgtgtg tttctcagga gaccgggggg gtgtcttagc acttgctgtg gtgtagatgg
     1501 aggactctgg gctcagcaca gccttgtcca aaagcgtggc acactccagt ccagcgacag
     1561 cagccgcagc actcccccgg ttcctgagag ggtcatggta catccctaga acggggagga
     1621 gtggactggg gtcaccctcc atgaagttga ctttaagtgc tcggagcttc aaagggttgt
     1681 tagttttcat ggaactcttg gggctctcga tgaagaaact agagtgctgg ctggcatcag
     1741 ggctggggtt ggcctccaca aacctaccac cactggcacc cagagctccc cgccagccca
     1801 cttctggggc tgtgctgccc ccagtcttgc tggggagtga tgtcaaaggg ctgtgggaga
     1861 atctggtggc ctcagggaag tctgtggcac agctaatgaa actatcaatg cttgacatac
     1921 ttcgcatgtc aatgacccct tctgtgcgag tgggcagagg ggctacgggg ctggggatac
     1981 tctccatttc aagttcttcc tttggtttgc tctgtgctgc tgactccttg gtattgagga
     2041 tgggttggga ttgggtcttg gccatcttat tgtacatgtc ttccaactgc tgaacgttca
     2101 ggtgctgagg actagaagac gatctggctt tttcagggga tccctgttcc ttggtttcaa
     2161 aggacttact tgagctggtt tcagacagtg tcctctttgt ccatttccat ttgttaggag
     2221 acaagtggtt atcttgtact ttgtctttct tacccatatt ctccccattt ttctcaacca
     2281 caatgtccat catctcaatg ctccgggcaa aagcatcctt catgttcatg gatacgatgc
     2341 tgccattcct cttggctctc tccagagcct ctcgccgttt gattgctttc tcctgtctct
     2401 tctgctcctt atagaactca gagaagttat tgacgatgat ggggatggga agagcaatca
     2461 ccaggactcc tgcaatgcag cagagtcccc caacaatttt ccccaggaga gtcttggggt
     2521 agatgtctcc atacccaaca gtagtcatgg tgatggtggc ccaccagaaa gaggctggga
     2581 tgcttttgaa cttggtgtcg tcctcatcct tctcagcaaa gaagacaagg ctggagaaga
     2641 tcataatgcc catggcaagg aagaggatga gcaagcccaa ctcattgtag ctcctccgca
     2701 aagtgaagcc cagagactgg aggccagtgg agtggcgtgc aagcttaagg atgcggagaa
     2761 tgcgcatgat gcggaagatc tggaccacgc ggcggacatt ctggaattgc agcacgctct
     2821 tgttggattc ggtgaggaaa atggtgacat agtatggcag aatggccaac aagtcaatgg
     2881 cattgagtgg gcccttgaag aacttccact tcttgggcga ggagaggaac ctcagcaggt
     2941 actccatggt gaaccatgcg atgcacacgg cctccacgtg ggccagctgg gggttgtctg
     3001 tggactggcc gaactcatcg aggctctgta gctcaggcag cgtgttgagg gacagggcaa
     3061 tggtggagag gacgatgaac atgatggaaa ttatggcaag gatcttggca gccacagagg
     3121 aattgggctt ctccagtagg tcccagagtt ttttcctctt ctctgcgcag cacgtgttat
     3181 cgaactcctc gccttcccgc tcccgtaggg tctcggcctc acgcttgagc tcctcgttca
     3241 tctgctcttt cttctggtgg tagcgggcct ggcagcagga ctccgggtag atctcgtcga
     3301 tgccccagta gtcgagctct tggctgaagc tgagcgcgca catctcctcc atcatgtgca
     3361 gtcgcccagt gcggtagaag ttgaggatgg aggtgaaggc gcccgggtgg cggtcaaaga
     3421 agtactcgtt gtcgtcgagg ctgtagtcat cgcacacctc gagcagcgag tcgtgcgtgt
     3481 tgcagtcgcg gagcttgccc agccgcgtgc ggggcaggcg gtccagggta cgccagagta
     3541 cctcgtgcgc cagccccccg acgttgaggc ggacccgccg agagcacgcc ttgctgcgca
     3601 cgatctccat gggctcgggc ggcagcgagc tggtggagcg ggagccatgc ttcgtcatgc
     3661 ccgccggcat ttgaattcga agcttgagct cgagatctga gtccggactt gtacagctcg
     3721 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     3781 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     3841 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     3901 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     3961 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     4021 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     4081 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     4141 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     4201 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     4261 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     4321 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     4381 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc gaccggtagc
     4441 gctagcggat ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac
     4501 gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt
     4561 gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc
     4621 gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa
     4681 tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact
     4741 gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca
     4801 tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac
     4861 gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg
     4921 ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga
     4981 actccatata tgggctatga actaatgacc ccgtaattga ttactattaa taactaatgc
     5041 atggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5101 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5161 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5221 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5281 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5341 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5401 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5461 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5521 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5581 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5641 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5701 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5761 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5821 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5881 agtatatatg agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc
     5941 cctgggcctt cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat
     6001 tggccccaat ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg
     6061 cgtttatgaa caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata
     6121 gcgcgggttc cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg
     6181 gcgaagaact ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa
     6241 acgattccga agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc
     6301 aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt
     6361 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     6421 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     6481 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     6541 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     6601 cgtcgggcat gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     6661 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     6721 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     6781 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     6841 cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     6901 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt
     6961 gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     7021 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     7081 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     7141 tgcgaaacga tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa
     7201 aagcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
     7261 ataaaaaaaa ttagtcagcc atggggcgga gaatgggcg
//
