LOCUS       sequence-12987-e        8139 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 8139)
  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 8139)
  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: pCDNA3.1(+) GFP-PIPKI{gamma}661-FLAG
FEATURES             Location/Qualifiers
     source          1..8139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      935..944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(941..943,944..946,947..1657)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(941..943,944..946,947..1657)
                     /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(986..1007)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1247..1266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1594..1615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3665..3688
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3729..3746)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3735..3959
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4005..4433
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4092..4111)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4302..4323
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4442..4462)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4447..4777
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4628..4763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4690..4709
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4844..5638
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4898..4917)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5508..5527
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5812..5933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5849..5868)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5903..5922
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5982..5998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5982..5998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5995..6017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6006..6022
                     /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)."
     promoter        complement(join(6030..6036,6037..6054,6055..6060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6075..6096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6213..6230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6384..6972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6464..6483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7143..7934,7935..8003))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7766..7785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8004..8108)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atggtgagca agggcgagga
      961 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1021 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1081 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1141 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1201 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     1261 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     1321 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     1381 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     1441 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     1501 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     1561 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1621 cgccgggatc actctcggca tggacgagct gtacaagtcc ggactcagat ccatggagct
     1681 agaggtgccg gacgaggcgg agagcgccga agcgggggcc gtgacggcgg aagcggcctg
     1741 gtctgcggag agtggggcgg cggcaggtat gacccagaag aaggctggcc ttgcagaggc
     1801 ccccttggtg acagggcagc caggccctgg ccatgggaag aagctgggcc accgaggcgt
     1861 ggatgcgtcg ggagagacta cgtataagaa gaccacctcg tccaccctga agggcgccat
     1921 ccagctgggg atcgggtaca cggtgggcaa cctgagctct aagccggagc gggacgtgct
     1981 catgcaggac ttctacgtga tggagagcat cttcttcccc agtgaaggga gcaacttcac
     2041 ccctgcccac cacttccagg atttccgctt caagacctat gcacctgttg ccttccgcta
     2101 cttccgggag ctctttggca tccgtccaga tgactatttg tactccctgt gcaatgagcc
     2161 actcatcgag ctctccaacc ccggggccag cggctctgtc ttctacgtca ccagcgacga
     2221 cgagttcatc atcaagactg tcatgcacaa ggaggcagag ttcctgcaga agctgctgcc
     2281 tggctactac atgaatctca accagaaccc acggacgctg ctgcccaagt tctatgggct
     2341 gtactgcgtg cagtctggtg gcaagaacat ccgcgtggtg gtcatgaaca atgtgctgcc
     2401 ccgtgtcgtc aaaatgcacc ttaagttcga cctcaagggc tccacgtaca agcgcagggc
     2461 cagcaagaag gagaaggaga agagcctgcc cacctacaag gacctggact tcatgcagga
     2521 catgcccgag gggctgctgc tggactccga cacctttggc gccctggtca agacgctgca
     2581 gcgagactgc ctggtgctgg agagcttcaa gataatggac tacagcctgc tgctgggcgt
     2641 gcacaacatc gatcagcagg agcgagagcg ccaggccgag ggcgcccaga gcaaggcgga
     2701 tgagaagcgg cccgtggccc agaaggctct gtattccacg gccatggagt ctatccaggg
     2761 cggagctgcc cgtggggagg ccattgagac agatgacacg atgggtggga ttccagcagt
     2821 gaatgggcgc ggggagcgac tgctcctcca catcgggatc attgatattc tgcagtccta
     2881 caggttcatc aagaagttag aacacacctg gaaggccctc gtccatgatg gggacactgt
     2941 ctcagtccac cggcccagct tctatgcaga gcgcttcttc aagttcatga gcagcacggt
     3001 gttccggaag agttcctccc tgaagtcctc tccatccaag aaaggacgtg gtgccctgct
     3061 ggcggtcaaa cccctggggc ccactgctgc cttctcagcc agccagatcc ccagcgagag
     3121 agaggatgtg cagtatgacc tgcggggggc ccgcagctac cccacgcttg aggatgaagg
     3181 ccggcctgac ctcctgccct gcaccccacc gtcctttgag gaagccacca cagcctccat
     3241 cgccaccacc ctgtcgtcca cctccctctc catcccagag cggtcccctt cagatacatc
     3301 ggagcagccc cggtacaggc ggcgcacgca gtcttcaggc caggatggcc ggccccagga
     3361 ggagccccat gcggaagacc tgcagaagat aactgtgcag gtggagccag tgtgcggtgt
     3421 gggggttgtc cccaaggagg agggtgcagg agtggaggtc cccccatgtg gggcatcggc
     3481 tgcagcctct gtggaaatag acgctgccag ccaggcctca gagcctgcca gccaggcctc
     3541 agatgaggag gatgcaccct ctacagacat ctattttccc accgacgaga ggagctgggt
     3601 gtactccccg cttcactata gcgcgcggcc cgcctccgac ggcgagagcg acacactcga
     3661 gatggactac aaggacgacg atgacaagaa gctttgatct agagggcccg tttaaacccg
     3721 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3781 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3841 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3901 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3961 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4021 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4081 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4141 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     4201 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4261 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4321 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     4381 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4441 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat
     4501 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4561 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4621 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4681 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     4741 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     4801 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     4861 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4921 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4981 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5041 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5101 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5161 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5221 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5281 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5341 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5401 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5461 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5521 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5581 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5641 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5701 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5761 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5821 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5881 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5941 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     6001 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6061 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6121 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6181 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6241 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6301 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6361 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6421 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6481 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6541 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     6601 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6661 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6721 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6781 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     6841 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6901 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6961 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     7021 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7081 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7141 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7201 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7261 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7321 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7381 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7441 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7501 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7561 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7621 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7681 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7741 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7801 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7861 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7921 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7981 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     8041 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     8101 gttccgcgca catttccccg aaaagtgcca cctgacgtc
//
