LOCUS       sequence-456541-        6933 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 6933)
  AUTHORS   Song Y, Maul RS, Gerbin CS, Chang DD
  TITLE     Inhibition of anchorage-independent growth of transformed NIH3T3 
            cells by epithelial protein lost in neoplasm (EPLIN) requires 
            localization of EPLIN to actin cytoskeleton.
  JOURNAL   Mol Biol Cell. 2002 Apr;13(4):1408-16.
  PUBMED    11950948
REFERENCE   2  (bases 1 to 6933)
  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."; date: "2002-04"; volume: "13(4)"; pages: "1408-16"
COMMENT     Sequence Label: pCMV5-Flag-EPLIN beta
FEATURES             Location/Qualifiers
     source          1..6933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..62
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     54..71
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     55..71
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        232..611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        612..815
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     766..786
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      838..847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             847..870
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3189..3811
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3388..3407)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3835..3855)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3840..4169
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4020..4155
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4207..4226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4208..4226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4240..4256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4240..4256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4253..4275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4264..4280
                     /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(4288..4294,4295..4312,4313..4318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4333..4354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4471..4488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4642..5229)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4722..4741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5400..6191,6192..6260))
                     /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     6023..6042
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6261..6365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6392..6847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6479..6498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6689..6710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
       61 cgacggccag tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag
      121 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
      181 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
      241 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      481 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      541 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
      601 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
      661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      721 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
      781 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagaattgcc
      841 accatggact acaaggacga cgatgacaag aattctgcag gatccaagat ggaatcatct
      901 ccatttaata gacggcaatg gacctcacta tcattgaggg taacagccaa agaactttct
      961 cttgtcaaca agaacaagtc atcggctatt gtggaaatat tctccaagta ccagaaagca
     1021 gctgaagaaa caaacatgga gaagaagaga agtaacaccg aaaatctctc ccagcacttt
     1081 agaaagggga ccctgactgt gttaaagaag aagtgggaga acccagggct gggagcagag
     1141 tctcacacag actctctacg gaacagcagc actgagatta ggcacagagc agaccatcct
     1201 cctgctgaag tgacaagcca cgctgcttct ggagccaaag ctgaccaaga agaacaaatc
     1261 caccccagat ctagactcag gtcacctcct gaagccctcg ttcagggtcg atatccccac
     1321 atcaaggacg gtgaggatct taaagaccac tcaacagaaa gtaaaaaaat ggaaaattgt
     1381 ctaggagaat ccaggcatga agtagaaaaa tcagaaatca gtgaaaacac agatgcttcg
     1441 ggcaaaatag agaaatataa tgttccgctg aacaggctta agatgatgtt tgagaaaggt
     1501 gaaccaactc aaactaagat tctccgggcc caaagccgaa gtgcaagtgg aaggaagatc
     1561 tctgaaaaca gctattctct agatgacctg gaaataggcc caggtcagtt gtcatcttct
     1621 acatttgact cggagaaaaa tgagagtaga cgaaatctgg aacttccacg cctctcagaa
     1681 acctctataa aggatcgaat ggccaagtac caggcagctg tgtccaaaca aagcagctca
     1741 accaactata caaatgagct gaaagccagt ggtggcgaaa tcaaaattca taaaatggag
     1801 caaaaggaga atgtgccccc aggtcctgag gtctgcatca cccatcagga aggggaaaag
     1861 atttctgcaa atgagaatag cctggcagtc cgttccaccc ctgccgaaga tgactcccgt
     1921 gactcccagg ttaagagtga ggttcaacag cctgtccatc ccaagccact aagtccagat
     1981 tccagagcct ccagtctttc tgaaagttct cctcccaaag caatgaagaa gtttcaggca
     2041 cctgcaagag agacctgcgt ggaatgtcag aagacagtct atccaatgga gcgtctcttg
     2101 gccaaccagc aggtgtttca catcagctgc ttccgttgct cctattgcaa caacaaactc
     2161 agtctaggaa catatgcatc tttacatgga agaatctatt gtaagcctca cttcaatcaa
     2221 ctctttaaat ctaagggcaa ctatgatgaa ggctttgggc acagaccaca caaggatcta
     2281 tgggcaagca aaaatgaaaa cgaagagatt ttggagagac cagcccagct tgcaaatgca
     2341 agggagaccc ctcacagccc aggggtagaa gatgccccta ttgctaaggt gggtgtcctg
     2401 gctgcaagta tggaagccaa ggcctcctct cagcaggaga aggaagacaa gccagctgaa
     2461 accaagaagc tgaggatcgc ctggccaccc cccactgaac ttggaagttc aggaagtgcc
     2521 ttggaggaag ggatcaaaat gtcaaagccc aaatggcctc ctgaagacga aatcagcaag
     2581 cccgaagttc ctgaggatgt cgatctagat ctgaagaagc taagacgatc ttcttcactg
     2641 aaggaaagaa gccgcccatt cactgtagca gcttcatttc aaagcacctc tgtcaagagc
     2701 ccaaaaactg tgtccccacc tatcaggaaa ggctggagca tgtcagagca gagtgaagag
     2761 tctgtgggtg gaagagttgc agaaaggaaa caagtggaaa atgccaaggc ttctaagaag
     2821 aatgggaatg tgggaaaaac aacctggcaa aacaaagaat ctaaaggaga gacagggaag
     2881 agaagtaagg aaggtcatag tttggagatg gagaatgaga atcttgtaga aaatggtgca
     2941 gactccgatg aagatgataa cagcttcctc aaacaacaat ctccacaaga acccaagtct
     3001 ctgaattggt cgagttttgt agacaacacc tttgctgaag aattcactac tcagaatcag
     3061 aaatcccagg atgtggaact ctgggaggga gaagtggtca aagagctctc tgtggaagaa
     3121 cagataaaga gaaatcggta ttatgatgag gatgaggatg aagagtgagt cgacatcgat
     3181 taaagcttgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg
     3241 ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact
     3301 aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg
     3361 ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac
     3421 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     3481 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     3541 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     3601 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3661 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     3721 ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     3781 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg
     3841 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3901 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3961 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4021 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4081 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4141 gcttttttgg aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa aaccagaaag
     4201 ttaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     4261 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4321 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4381 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4441 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4501 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4561 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4621 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4681 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4741 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4801 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4861 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4921 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4981 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5041 agagttcttg aagtgtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5101 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5161 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5221 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5281 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5341 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5401 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5461 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5521 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5581 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5641 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5701 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5761 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5821 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5881 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5941 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6001 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6061 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6121 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6181 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6241 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6301 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6361 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     6421 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     6481 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     6541 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     6601 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6661 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6721 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6781 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     6841 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     6901 cttcgctatt acgccagctg gcgaaagggg gat
//
