LOCUS       sequence-48796-e        6082 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 6082)
  AUTHORS   Pellegrin S, Mellor H
  TITLE     The Rho family GTPase Rif induces filopodia through mDia2.
  JOURNAL   Curr Biol. 2005 Jan 26;15(2):129-33.
  PUBMED    15668168
REFERENCE   2  (bases 1 to 6082)
  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: "Curr 
            Biol."; date: "2005-01-26"; volume: "15(2)"; pages: "129-33"
COMMENT     Sequence Label: myc Rif-TN
FEATURES             Location/Qualifiers
     source          1..6082
                     /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"
     CDS             909..938
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1608..1624)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1608..1624)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(1634..1652)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1635..1652)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1672..1689)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1678..1902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1948..2376
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2035..2054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2245..2266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2385..2405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2390..2720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2571..2706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2633..2652
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2787..3581
                     /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(2841..2860)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3451..3470
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3755..3876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3792..3811)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3846..3865
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3925..3941)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3925..3941)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3938..3960)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3949..3965
                     /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(3973..3979,3980..3997,3998..4003))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4018..4039
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4156..4173)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4327..4915)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4407..4426)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5086..5877,5878..5946))
                     /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     5709..5728
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5947..6051)
                     /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 gcttgatagc
      901 ttaccatgga acaaaaacta atatcggaag aagatctagg gcccatggat gcccccggag
      961 ccctggccca gaccgccgcc cccggtccgg gcaggaagga gctgaagatc gtgatcgtgg
     1021 gcgacggcgg ctgcggcaag aactcgctgc tcatggtgta cagccagggc tccttccccg
     1081 agcactacgc cccatcggtg ttcgagaagt acacggccag cgtgaccgtt ggcagcaagg
     1141 aggtgaccct gaacctctac gacacggccg ggcaagaaga ctatgaccgg ctgcggcccc
     1201 tgtcctacca gaacacccac ctcgtgctca tctgctatga cgtcatgaat cccaccagct
     1261 acgacaacgt cctcatcaag tggttccctg aggtcacgca tttctgccgc gggatcccca
     1321 tggtgctcat cggctgcaag acagacctga ggaaggacaa ggagcagctg cggaagctcc
     1381 gggccgccca gctggagccc atcacctaca tgcagggcct gagcgcctgc gaacagatcc
     1441 gagctgctct ctacctggaa tgttccgcca agtttcggga gaatgtggag gacgtcttcc
     1501 gggaggccgc caaggtggct ctcagcgctc tgaagaaggc gcaacggcag aagaagcgcc
     1561 ggctctgcct gctgctctga tctagatcga attcctgcag cccgggggat ccactagttc
     1621 tagagggccg tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     1681 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     1741 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     1801 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     1861 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     1921 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     1981 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2041 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2101 ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2161 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2221 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2281 ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa
     2341 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     2401 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2521 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2581 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2641 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2701 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     2761 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2821 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2881 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2941 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3001 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3061 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3121 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3181 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3241 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3301 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3361 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3421 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3481 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3541 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3601 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3661 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3721 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3781 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3841 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3901 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4501 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6061 ccgaaaagtg ccacctgacg tc
//
