LOCUS       sequence-482206-        4197 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Mammalian expression empty backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4197)
  TITLE     Vectors
REFERENCE   2  (bases 1 to 4197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: 1435 pSG5L Flag HA
FEATURES             Location/Qualifiers
     source          1..4197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..211
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        23..219
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      70..205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          250..822
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     830..849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     877..896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        877..895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             926..949
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             962..988
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             962..988
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1059..1180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1096..1115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1150..1169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1214..1231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1385..1973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1465..1484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2144..2935,2936..3004))
                     /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     2767..2786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3005..3109)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3177..3195
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3233..3255)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3355..3374
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3441..3896)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3578..3599)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3790..3809
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4022..4044
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4036..4053
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4037..4053
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4090..4110)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
       61 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      121 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      181 tagtgaagag gcttttttgg aggcctaggc ttttgcaaaa agctccggat cgatcctgag
      241 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
      301 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
      361 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
      421 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
      481 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
      541 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
      601 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
      661 ggaaatattc ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta
      721 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
      781 ccctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
      841 gttattgtgc tgtctcatca ttttggcaaa gaattgtaat acgactcact atagggcgaa
      901 ttgtcgaccc gggaagctta ccatggacta caaggacgac gatgacaagc tcgatggagg
      961 atacccctac gacgtgcccg actacgccag cggatccgcg cggatccgcg aattccccga
     1021 tatcgcctcg agtctagatc cagatcttat taaagcagaa cttgtttatt gcagcttata
     1081 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1141 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg tcgactctag
     1201 actcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     1261 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     1321 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     1381 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     1441 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     1501 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     1561 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     1621 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     1681 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     1741 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     1801 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     1861 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     1921 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     1981 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2161 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     2221 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     2281 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     2341 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     2401 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     2461 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     2521 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     2581 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     2641 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     2701 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     2761 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     2821 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     2881 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     2941 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     3001 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     3061 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     3121 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     3181 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     3241 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     3301 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     3361 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     3421 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     3481 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     3541 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     3601 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     3661 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     3721 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     3781 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     3841 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat
     3901 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     3961 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     4021 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt cgacttctga ggcggaaaga
     4081 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4141 gaagtatgca aagcatgcat ctcaattagt cagcaaggaa agtccccagg ctcccca
//
