LOCUS       Exported                6221 bp ds-DNA     circular SYN 05-MAY-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6221)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6221
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..589
                     /label=CMV
                     /note="CMV"
     misc_feature    1
                     /label=GFP tag
                     /note="GFP tag"
     misc_feature    1
                     /label=Kanamycin
                     /note="Kanamycin"
     primer_bind     complement(592..630)
                     /label=iRFP670_C1Swap_R
     regulatory      607..616
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1557
                     /codon_start=1
                     /product="bright monomeric near-infrared fluorescent 
                     protein engineered from a bacterial phytochrome, with an 
                     emission maximum of 670 nm (Shcherbakova et al., 2016)"
                     /label=miRFP670
                     /label=ORF frame 1
                     /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH
                     RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE
                     YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG
                     YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT
                     YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV
                     VCHHYLPRFIRFELRAICKRLAERIATRITALES"
     primer_bind     613..636
                     /label=iRFP670_F
     CDS             613..616
                     /codon_start=1
                     /label=iRFP
                     /translation="M"
     primer_bind     complement(1537..1558)
                     /label=piRFP670_R
     primer_bind     complement(1537..1557)
                     /label=piRFP670NEW_R
     primer_bind     1540..1578
                     /label=iRFP670_C1SwapNEW_F
     primer_bind     1543..1578
                     /label=iRFP670_C1Swap_F
     misc_feature    1558..1580
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     complement(1558..1577)
                     /label=RepIRFPr
     primer_bind     1558..1577
                     /label=iRFP-Fis1-vF
     misc_feature    1581..1611
                     /label=MCS
                     /note="MCS"
     CDS             1612..2877
                     /codon_start=1
                     /gene="PIP4K2C"
                     /gene_synonym="PIP5K2C"
                     /product="phosphatidylinositol 5-phosphate 4-kinase type-2
                     gamma isoform a"
                     /EC_number="2.7.1.149"
                     /label=PIP4K2C
                     /note="isoform a is encoded by transcript variant 1; 
                     phosphatidylinositol-4-phosphate 5-kinase, type II, gamma; 
                     phosphatidylinositol 4-phosphate 5-kinase; PIP4KII-gamma; 
                     PI(5)P 4-kinase type II gamma; phosphatidylinositol 
                     5-phosphate 4-kinase type II gamma"
                     /db_xref="CCDS:CCDS8946.1"
                     /db_xref="GeneID:79837"
                     /db_xref="HGNC:HGNC:23786"
                     /db_xref="MIM:617104"
                     /protein_id="NP_079055.3"
                     /translation="MASSSVPPATVSAATAGPGPGFGFASKTKKKHFVQQKVKVFRAAD
                     PLVGVFLWGVAHSINELSQVPPPVMLLPDDFKASSKIKVNNHLFHRENLPSHFKFKEYC
                     PQVFRNLRDRFGIDDQDYLVSLTRNPPSESEGSDGRFLISYDRTLVIKEVSSEDIADMH
                     SNLSNYHQYIVKCHGNTLLPQFLGMYRVSVDNEDSYMLVMRNMFSHRLPVHRKYDLKGS
                     LVSREASDKEKVKELPTLKDMDFLNKNQKVYIGEEEKKIFLEKLKRDVEFLVQLKIMDY
                     SLLLGIHDIIRGSEPEEEAPVREDESEVDGDCSLTGPPALVGSYGTSPEGIGGYIHSHR
                     PLGPGEFESFIDVYAIRSAEGAPQKEVYFMGLIDILTQYDAKKKAAHAAKTVKHGAGAE
                     ISTVHPEQYAKRFLDFITNIFA"
     primer_bind     1612..1634
                     /label=PIP4K2C_KpnICut_F
     primer_bind     complement(2424..2448)
                     /label=PIP4K2C_D280_R
     CDS             2449..2451
                     /codon_start=1
                     /label=D280
                     /translation="D"
     primer_bind     2452..2475
                     /label=PIP4K2C_D280K_F
     primer_bind     complement(2850..2877)
                     /label=PIP4K2C_BamHICut_R
     misc_feature    2878..2916
                     /label=MCS
                     /note="MCS"
     primer_bind     complement(2915..2934)
                     /label=SV40-pArev
     polyA_signal    3009..3130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    3137..3592
                     /label=f1
                     /note="f1"
     promoter        3619..3723
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3654..3682
                     /label=Bacterial
                     /note="Bacterial"
     promoter        3725..4082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        3766..3995
                     /label=SV40early
                     /note="SV40early"
     misc_feature    3933..4068
                     /label=SV40
                     /note="SV40"
     misc_feature    4117..4911
                     /label=Kan/Neo
                     /note="Kan/Neo"
     polyA_signal    5143..5190
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    5147..5165
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    5496..6139
                     /label=pUC
                     /note="pUC"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtagc aggtcatgcc tctggcagcc ccgcattcgg gaccgcctct
      661 cattcgaatt gcgaacatga agagatccac ctcgccggct cgatccagcc gcatggcgcg
      721 cttctggtcg tcagcgaaca tgatcatcgc gtcatccagg ccagcgccaa cgccgcggaa
      781 tttctgaatc tcggaagcgt actcggcgtt ccgctcgccg agatcgacgg cgatctgttg
      841 atcaagatcc tgccgcatct cgatcccacc gccgaaggca tgccggtcgc ggtgcgctgc
      901 cggatcggca atccctctac ggagtactgc ggtctgatgc atcggcctcc ggaaggcggg
      961 ctgatcatcg aactcgaacg tgccggcccg tcgatcgatc tgtcaggcac gctggcgccg
     1021 gcgctggagc ggatccgcac ggcgggttca ctgcgcgcgc tgtgcgatga caccgtgctg
     1081 ctgtttcagc agtgcaccgg ctacgaccgg gtgatggtgt atcgtttcga tgagcaaggc
     1141 cacggcctgg tattctccga gtgccatgtg cctgggctcg aatcctattt cggcaaccgc
     1201 tatccgtcgt cgactgtccc gcagatggcg cggcagctgt acgtgcggca gcgcgtccgc
     1261 gtgctggtcg acgtcaccta tcagccggtg ccgctggagc cgcggctgtc gccgctgacc
     1321 gggcgcgatc tcgacatgtc gggctgcttc ctgcgctcga tgtcgccgtg ccatctgcag
     1381 ttcctgaagg acatgggcgt gcgcgccacc ctggcggtgt cgctggtggt cggcggcaag
     1441 ctgtggggcc tggttgtctg tcaccattat ctgccgcgct tcatccgttt cgagctgcgg
     1501 gcgatctgca aacggctcgc cgaaaggatc gcgacgcgga tcaccgcgct tgagagctcc
     1561 ggactcagat ctcgagctca agcttcgaat tctgcagtcg acggtaccgc gatggcgtcc
     1621 tcctcggtcc caccagccac ggtatcggcg gcgacagcag gccccggccc aggtttcggc
     1681 ttcgcctcca agaccaagaa gaagcatttc gtgcagcaga aggtgaaggt gttccgggcg
     1741 gccgacccgc tggtgggtgt gttcctgtgg ggcgtagccc actcgatcaa tgagctcagc
     1801 caggtgcctc ccccggtgat gctgctgcca gatgacttta aggccagctc caagatcaag
     1861 gtcaacaatc accttttcca cagggaaaat ctgcccagtc atttcaagtt caaggagtat
     1921 tgtccccagg tcttcaggaa cctccgtgat cgatttggca ttgatgacca agattacttg
     1981 gtgtccctta cccgaaaccc ccccagcgaa agtgaaggca gtgatggtcg cttccttatc
     2041 tcctacgatc ggactctggt catcaaagaa gtatccagtg aggacattgc tgacatgcat
     2101 agcaacctct ccaactatca ccagtacatt gtgaagtgcc atggcaacac gcttctgccc
     2161 cagttcctgg ggatgtaccg agtcagtgtg gacaacgaag acagctacat gcttgtgatg
     2221 cgcaatatgt ttagccaccg tcttcctgtg cacaggaagt atgacctcaa gggttcccta
     2281 gtgtcccggg aagccagcga taaggaaaag gttaaagaat tgcccaccct taaggatatg
     2341 gactttctca acaagaacca gaaagtatat attggtgaag aggagaagaa aatatttctg
     2401 gagaagctga agagagatgt ggagtttcta gtgcagctga agatcatgga ctacagcctt
     2461 ctgctaggca tccacgacat cattcggggc tctgaaccag aggaggaagc gcccgtgcgg
     2521 gaggatgagt cagaggtgga tggggactgc agcctgactg gacctcctgc tctggtgggc
     2581 tcctatggca cctccccaga gggtatcgga ggctacatcc attcccatcg gcccctgggc
     2641 ccaggagagt ttgagtcctt cattgatgtc tatgccatcc ggagtgctga aggagccccc
     2701 cagaaggagg tctacttcat gggcctcatt gatatcctta cacagtatga tgccaagaag
     2761 aaagcagctc atgcagccaa aactgtcaag catggggctg gggcagagat ctctactgtc
     2821 catccggagc agtatgctaa gcgattcctg gattttatta ccaacatctt tgcctaacgg
     2881 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2941 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3001 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3061 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3121 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3181 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3241 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3301 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3361 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3421 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3481 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3541 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3601 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3661 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3721 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3781 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3841 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3901 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4021 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4081 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4141 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4201 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4261 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4321 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4381 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4441 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4501 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4561 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4621 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4681 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4741 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4801 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4861 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4921 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4981 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5041 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     5101 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5161 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5221 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5281 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5341 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5401 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5461 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5521 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5581 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5641 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5701 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5761 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5821 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5881 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5941 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     6001 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     6061 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6121 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6181 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
