LOCUS       sequence-311856-        7708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian cell expression of mouse LIG3-179-936.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7708)
  TITLE     Jianjun Zhao Lab Plasmids
REFERENCE   2  (bases 1 to 7708)
  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"
COMMENT     Sequence Label: pcDNA3-3NFLAG-mLIG3-179-936
FEATURES             Location/Qualifiers
     source          1..7708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        175..554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        555..758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     709..729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     803..822
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        803..821
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             844..867
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             871..894
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             898..921
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1214..1223
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3239..3256)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3245..3469
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3515..3943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3602..3621)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3812..3833
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3952..3972)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3957..4286
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4137..4272
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4199..4218
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4353..5147
                     /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(4407..4426)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5017..5036
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5321..5442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5358..5377)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5412..5431
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5491..5507)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5491..5507)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5504..5526)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5515..5531
                     /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(5539..5545,5546..5563,5564..5569))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5584..5605
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5722..5739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5893..6481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5973..5992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6652..7443,7444..7512))
                     /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     7275..7294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7513..7617)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7692..7708,1..3))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
       61 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      121 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      181 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      541 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      721 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      781 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      841 atggattaca aggacgatga cgataagggt gattacaagg acgatgacga taagggtgat
      901 tacaaggacg atgacgataa gcttggtacc agccagcata ttgcagacct gtcttctaag
      961 gcagctgcca cacctaagaa gaaagctgct gtccaggcta agctgacaac cactggccag
     1021 gtgacatctc cagtgaaagg tgcttcgttt attaccagta ccaatcctcg gaagttctct
     1081 ggcttttcag cagcaaaacc caacaactct gagcaagccc cctcaagccc tgctcctgga
     1141 acgagtctgt ctgcaagtaa atgtgaccct aagcacaaag attgtctgct acgagagttc
     1201 cggaagctgt gcgccatggt ggctgaaaat cctagctata acaccaagac ccagatcatc
     1261 catgacttct tgcagaaagg ctccacagga gatggctttc acggtgatgt gtacctgaca
     1321 gttaagctgc tgctgcctgg tgtcatcaag agtgtttaca acttgaatga taagcagatt
     1381 gtaaaacttt ttagccgaat ttttaactgc aacccagatg atatggcccg ggacctagaa
     1441 cagggtgacg tgtcagagac gatcagaatc ttctttgagc agagcaagtc tttcccccca
     1501 gctgccaaaa gcctcctcac cattcaggaa gtggatgcct tcctcctgca cctctccaag
     1561 ctcaccaaag aggatgagca gcagcaggcc ctgcaggaca tagcctcccg gtgtacagcc
     1621 aatgacctta agtgcattat tcggctgatc aaacatgatc tgaagatgaa ctcgggtgca
     1681 aaacacgtgt tagatgccct tgaccccaat gcttatgaag ccttcaaagc atcgcgcaac
     1741 ctgcaggatg tggtggagcg agtccttcac aatgagcagg aggtggagaa ggacccaggc
     1801 cggcgacggg ctctgagggt ccaggcctca ctgatgacgc ctgtgcagcc catgctggct
     1861 gaggcctgca aatccattga atatgcaatg aagaagtgtc ccaatggcat gttctctgag
     1921 atcaagtacg atggtgagcg agtccaggtg cataagaagg gggaccactt cagctacttc
     1981 agccgcagtc tcaagcctgt cctgcctcac aaggtggccc acttcaagga ctatataccc
     2041 aaagcttttc ctgggggtca aagcatgatc ttggattccg aggtactcct gattgacaac
     2101 aacacaggca aaccactgcc ctttgggact ctgggagtgc acaagaaagc tgctttccag
     2161 gatgctaatg tctgcctgtt tgtttttgat tgtatctact ttaatgatgt cagcttgatg
     2221 gacaggcctc tctgtgagcg acggaagttt cttcatgaca acatggttga aatccgtaac
     2281 cggatcatgt tctcagaaat gaagcaagtc acaaaagctt cagacctggc tgacatgata
     2341 aaccgggtga tccgagaggg cttggaaggg ttggtgctaa aggatgtgaa gggtacatat
     2401 gagcctggga agcggcactg gctgaaagtt aagaaagatt atttgaatga gggggccatg
     2461 gctgatacag ctgatctggt ggttcttggg gccttctatg ggcaagggag caaaggcgga
     2521 atgatgtcca tcttcctcat gggctgctat gaccctgaca gccaaaagtg gtgcactgtt
     2581 acaaaatgtg ctggaggcca tgatgatgcc acacttgccc gcctgcagaa agagctggtt
     2641 atggtgaaga tcagcaagga tcctagtaag atacccagct ggctgaagat caacaagatc
     2701 tattatcctg actttatcgt cccagaccca aagaaagctg ctgtgtggga gatcacaggg
     2761 gcagaattct ccagatctga ggcacacact gctgatggca tctccatccg atttcctcga
     2821 tgcacccgaa tccgggacga caaggactgg aaatctgcca ctaacctccc ccaactcaag
     2881 gaactgtacc agctgtccaa agacaaggca gactttgctg tggtggctgg agatgaggca
     2941 agccccacta ctggaggcag cagtggcgag aacgagggca ctgctgggtc tgctgggccc
     3001 tgcaagggcc cccccagtaa gtcctctgcc agtgccaaga ccaccgaaca gaagctcaat
     3061 agccccagta gcagaggtgg catcaaaccg attccaaagc attcccccat gaaaccagga
     3121 gagaagctgg ctgtgaagtc ttctccagtc aaagtgggaa tgaagaggaa agctactgat
     3181 gagaccccat gcctgaaaaa gtaataatct agagggcccg tttaaacccg ctgatcagcc
     3241 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3301 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3361 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3421 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3481 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3541 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3601 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3661 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3721 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3781 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3841 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3901 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3961 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4021 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4081 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4141 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4201 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4261 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4321 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4381 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4441 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4501 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4561 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4621 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4681 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4741 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4801 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4861 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4921 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4981 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5041 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5101 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5161 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5221 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5281 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     5341 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5401 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     5461 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     5521 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5581 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5641 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5701 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5761 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5821 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5881 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5941 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6001 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6061 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6121 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6181 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6241 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6301 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6361 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6421 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6601 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6661 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6721 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6781 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6841 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6901 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6961 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     7021 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     7081 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     7141 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7201 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7261 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7321 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7381 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7441 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7501 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7561 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7621 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     7681 tgcactctca gtacaatctg ctctgatg
//
