LOCUS       sequence-174746-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty LIC vector; adds a tdTomato gene to the C-terminus of your 
            protein of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  TITLE     6-series mammalian LIC expression vectors
REFERENCE   2  (bases 1 to 6858)
  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 tdTomato LIC cloning vector (6F)
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        188..567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        568..771
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     722..742
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     816..835
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        816..834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2364..2382)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2365..2382)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2402..2419)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2408..2632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2678..3106
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2765..2784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2975..2996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3115..3135)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3120..3449
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3300..3435
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3362..3381
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3516..4310
                     /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(3570..3589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4180..4199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4484..4605
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4521..4540)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4575..4594
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4654..4670)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4654..4670)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4667..4689)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4678..4694
                     /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(4702..4708,4709..4726,4727..4732))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4747..4768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4885..4902)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5056..5644)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5136..5155)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5815..6606,6607..6675))
                     /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     6438..6457
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6676..6780)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(6855..6858,1..16))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
       61 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      121 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      181 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      781 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      841 caagcttggt accgagctcg gatccctact tccaatccaa tattggtagt gggagcaacg
      901 gcagcagcgg atccgtgagt aagggcgagg aagtgatcaa agagttcatg cggtttaagg
      961 tgagaatgga aggaagcatg aacggccacg agttcgaaat tgagggagaa ggagagggac
     1021 ggccctacga gggcacccag acagccaagc tgaaagtgac aaagggcggg cctctgccat
     1081 tcgcttggga catcctgagc ccacagttta tgtacggctc caaggcctat gtgaaacatc
     1141 cagctgacat tcccgattat aagaaactga gcttccccga ggggtttaag tgggaaagag
     1201 tgatgaactt cgaggacgga ggcctggtga ctgtgaccca ggacagctcc ctgcaggatg
     1261 ggaccctgat ctacaaggtg aaaatgagag ggacaaattt tccccctgat ggacctgtga
     1321 tgcagaagaa aactatggga tgggaggcct ccaccgaaag gctgtatcca cgcgacgggg
     1381 tgctgaaagg agaaatccac caggctctga agctgaaaga tgggggacat tacctggtgg
     1441 agttcaagac aatctacatg gccaagaaac ctgtgcagct gccaggctac tattacgtgg
     1501 acacaaaact ggatatcact tcacacaacg aggactacac tattgtggag cagtatgaac
     1561 ggagcgaggg gagacaccat ctgttcctgg gccatgggac tggaagtacc ggctcagggt
     1621 ctagtggaac cgcctcaagc gaggataaca atatggctgt gatcaaagag ttcatgaggt
     1681 ttaaggtgcg catggagggc agcatgaatg ggcacgaatt tgagattgaa ggagagggcg
     1741 aagggaggcc ttacgagggc acacagactg ccaagctgaa agtgaccaag ggaggaccac
     1801 tgcctttcgc ttgggatatc ctgtctcctc agtttatgta cggaagtaag gcctatgtca
     1861 agcatcccgc tgacattcct gattacaaga aactgtcttt cccagagggc tttaagtggg
     1921 agagagtgat gaattttgaa gatggaggcc tggtgaccgt gacacaggac tcctctctgc
     1981 aggatggcac tctgatctac aaagtcaaaa tgcgcggcac caattttcca cccgatgggc
     2041 ccgtgatgca gaagaaaaca atggggtggg aggccagcac tgaacggctg tatcctagag
     2101 acggagtgct gaagggcgaa atccaccagg ccctgaagct gaaagacggc ggccactacc
     2161 tggtggagtt caaaaccatc tacatggcca agaaaccagt gcagctgccc ggctattact
     2221 atgtggacac caagctggat atcacatccc acaatgaaga ctacaccatt gtggaacagt
     2281 atgagaggtc tgaaggacgc caccatctgt ttctgtacgg catggatgag ctgtataagt
     2341 aatagtaatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2401 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2461 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2521 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2581 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2641 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2701 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2761 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2821 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2881 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2941 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3001 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3061 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3121 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3181 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3241 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3301 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3361 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3421 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3481 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3661 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4021 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     4081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4141 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4321 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4381 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4441 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     4501 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4561 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     4621 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4681 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4741 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4801 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4861 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4921 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4981 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5041 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5101 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5161 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5221 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5281 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5341 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5401 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5461 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5521 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5581 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5641 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5701 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5761 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5821 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5881 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5941 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6001 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6061 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6121 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6181 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6241 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6301 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6361 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6421 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6481 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6541 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6601 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6661 atgttgaata ctcatactct tcctttttca ttattattga agcatttatc agggttattg
     6721 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6781 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6841 tggtgcactc tcagtaca
//
