LOCUS       sequence-465731-        4260 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Allow Halotag(R) insert with weakened CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4260)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4260)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFC211A HaloTag-NLS CMVd1 Flexi Vector
FEATURES             Location/Qualifiers
     source          1..4260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..100
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     51..71
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     132..151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        156..174
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     156..173
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             213..548
                     /codon_start=1
                     /product="ribonuclease from Bacillus amyloliquefaciens"
                     /label=barnase
                     /note="The barnase gene is lethal in standard bacterial 
                     transformation strains."
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     CDS             588..608
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             624..1508
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             1518..1538
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1542..1562
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1566..1586
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1674..1692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1729..1850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1740..1759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1794..1813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2097..2201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2202..2270,2271..3062)
                     /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     complement(2420..2439)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3300..3319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3924..4207
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
       61 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      121 cactagaagc ttaatacgac tcactatagg gtacgattta ggtgacacta tagaataagg
      181 agcgatcgcc atggaataag taaggaatcc acatggcaca ggttatcaac acgtttgacg
      241 gggttgcgga ttatcttcag acatatcata agctacctga taattacatt acaaaatcag
      301 aagcacaagc cctcggctgg gtggcatcaa aagggaacct tgcagacgtc gctccgggga
      361 aaagcatcgg cggagacatc ttctcaaaca gggaaggcaa actcccgggc aaaagcggac
      421 gaacatggcg tgaagcggat attaactata catcaggctt cagaaattca gaccggattc
      481 tttactcaag cgactggctg atttacaaaa caacggacca ttatcagacc tttacaaaaa
      541 tcagataatg tttaatgacc ccgtgtcgag ctctcgagcc aaccactgag gatctgtact
      601 ttcagagcga taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg
      661 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
      721 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
      781 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
      841 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
      901 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
      961 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     1021 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     1081 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     1141 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     1201 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     1261 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     1321 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     1381 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     1441 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga
     1501 tttccggagc cggcgatcca aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg
     1561 tagatccaaa aaagaagaga aaggtataat agaattctag agtcgacctg caggcatgca
     1621 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     1681 gcaataacta gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg
     1741 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1801 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1861 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt
     1921 aaaacctcta caaatgtggt aaaatcgaat tctaatggat cctctttgcg cttgcgtttt
     1981 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     2041 tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     2101 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     2161 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2221 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2281 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2341 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2401 tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2461 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2521 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2581 tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa
     2641 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2701 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2761 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2821 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2881 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2941 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     3001 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     3061 aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3901 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     3961 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     4021 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     4081 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     4141 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     4201 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
