LOCUS       sequence-12988-e        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  AUTHORS   Lokuta MA, Senetar MA, Bennin DA, Nuzzi PA, Chan KT, Ott VL, 
            Huttenlocher A
  TITLE     Type Igamma PIP kinase is a novel uropod component that regulates 
            rear retraction during neutrophil chemotaxis.
  JOURNAL   Mol Biol Cell. 2007 Dec . 18(12):5069-80.
  PUBMED    17928408
REFERENCE   2  (bases 1 to 6659)
  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"; journalName: "Mol Biol 
            Cell. 2007 Dec . 18(12):5069-80."
COMMENT     Sequence Label: mCherryC1-PIPKI{gamma}661
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..1310
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(748..766)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1016..1035
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3447..3568
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3484..3503)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3538..3557
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3575..4030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3712..3733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3924..3943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4057..4161
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4163..4520
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4163..4354
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4186..4206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4371..4506
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4555..5349
                     /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             4555..5349
                     /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(4609..4628)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5219..5238
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5537..5556)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5581..5628
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5957..6545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6446..6465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gctagcgcca
      601 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      661 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      781 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      841 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1021 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1081 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1141 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1201 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1261 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag agatctatgg
     1321 agctagaggt gccggacgag gcggagagcg ccgaagcggg ggccgtgacg gcggaagcgg
     1381 cctggtctgc ggagagtggg gcggcggcag gtatgaccca gaagaaggct ggccttgcag
     1441 aggccccctt ggtgacaggg cagccaggcc ctggccatgg gaagaagctg ggccaccgag
     1501 gcgtggatgc gtcgggagag actacgtata agaagaccac ctcgtccacc ctgaagggcg
     1561 ccatccagct ggggatcggg tacacggtgg gcaacctgag ctctaagccg gagcgggacg
     1621 tgctcatgca ggacttctac gtgatggaga gcatcttctt ccccagtgaa gggagcaact
     1681 tcacccctgc ccaccacttc caggatttcc gcttcaagac ctatgcacct gttgccttcc
     1741 gctacttccg ggagctcttt ggcatccgtc cagatgacta tttgtactcc ctgtgcaatg
     1801 agccactcat cgagctctcc aaccccgggg ccagcggctc tgtcttctac gtcaccagcg
     1861 acgacgagtt catcatcaag actgtcatgc acaaggaggc agagttcctg cagaagctgc
     1921 tgcctggcta ctacatgaat ctcaaccaga acccacggac gctgctgccc aagttctatg
     1981 ggctgtactg cgtgcagtct ggtggcaaga acatccgcgt ggtggtcatg aacaatgtgc
     2041 tgccccgtgt cgtcaaaatg caccttaagt tcgacctcaa gggctccacg tacaagcgca
     2101 gggccagcaa gaaggagaag gagaagagcc tgcccaccta caaggacctg gacttcatgc
     2161 aggacatgcc cgaggggctg ctgctggact ccgacacctt tggcgccctg gtcaagacgc
     2221 tgcagcgaga ctgcctggtg ctggagagct tcaagataat ggactacagc ctgctgctgg
     2281 gcgtgcacaa catcgatcag caggagcgag agcgccaggc cgagggcgcc cagagcaagg
     2341 cggatgagaa gcggcccgtg gcccagaagg ctctgtattc cacggccatg gagtctatcc
     2401 agggcggagc tgcccgtggg gaggccattg agacagatga cacgatgggt gggattccag
     2461 cagtgaatgg gcgcggggag cgactgctcc tccacatcgg gatcattgat attctgcagt
     2521 cctacaggtt catcaagaag ttagaacaca cctggaaggc cctcgtccat gatggggaca
     2581 ctgtctcagt ccaccggccc agcttctatg cagagcgctt cttcaagttc atgagcagca
     2641 cggtgttccg gaagagttcc tccctgaagt cctctccatc caagaaagga cgtggtgccc
     2701 tgctggcggt caaacccctg gggcccactg ctgccttctc agccagccag atccccagcg
     2761 agagagagga tgtgcagtat gacctgcggg gggcccgcag ctaccccacg cttgaggatg
     2821 aaggccggcc tgacctcctg ccctgcaccc caccgtcctt tgaggaagcc accacagcct
     2881 ccatcgccac caccctgtcg tccacctccc tctccatccc agagcggtcc ccttcagata
     2941 catcggagca gccccggtac aggcggcgca cgcagtcttc aggccaggat ggccggcccc
     3001 aggaggagcc ccatgcggaa gacctgcaga agataactgt gcaggtggag ccagtgtgcg
     3061 gtgtgggggt tgtccccaag gaggagggtg caggagtgga ggtcccccca tgtggggcat
     3121 cggctgcagc ctctgtggaa atagacgctg ccagccaggc ctcagagcct gccagccagg
     3181 cctcagatga ggaggatgca ccctctacag acatctattt tcccaccgac gagaggagct
     3241 gggtgtactc cccgcttcac tatagcgcgc ggcccgcctc cgacggcgag agcgacacat
     3301 aaggtaccgc gggcccggga tccaccggat ctagataact gatcataatc agccatacca
     3361 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3421 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3481 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3541 gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa tattttgtta
     3601 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3661 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3721 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     3781 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3841 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3901 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3961 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4021 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4081 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4141 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4201 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4261 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4381 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4441 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4501 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4561 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4621 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4681 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     4741 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4801 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4861 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4921 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4981 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     5041 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     5101 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     5161 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     5221 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     5281 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5341 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5401 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5461 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5521 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     5581 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     5641 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     5701 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     5761 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     5821 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5881 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5941 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6001 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6061 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     6121 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6181 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6241 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6301 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6361 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6421 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6481 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6541 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6601 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
