LOCUS       sequence-470850-        4484 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Transfection backbone with secreted HiBiT protein and TK promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4484)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 4484)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFN229K secHiBiT TK-neo Flexi Vector
FEATURES             Location/Qualifiers
     source          1..4484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        7..758
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             982..1317
                     /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"
     primer_bind     complement(1432..1450)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1487..1608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1498..1517)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1552..1571
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        1853..2210
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1853..2044
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1876..1896)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2061..2196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2225..2272
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2304..3098
                     /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(2358..2377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2968..2987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3162..3210
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(3449..4037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3529..3548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4153..4436
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 gatctaaatg agtcttcgga cctcgcgggg gccgcttaag cggtggttag ggtttgtctg
       61 acgcgggggg agggggaagg aacgaaacac tctcattcgg aggcggctcg gggtttggtc
      121 ttggtggcca cgggcacgca gaagagcgcc gcgatcctct taagcacccc cccgccctcc
      181 gtggaggcgg gggtttggtc ggcgggtggt aactggcggg ccgctgactc gggcgggtcg
      241 cgcgccccag agtgtgacct tttcggtctg ctcgcagacc cccgggcggc gccgccgcgg
      301 cggcgacggg ctcgctgggt cctaggctcc atggggaccg tatacgtgga caggctctgg
      361 agcatccgca cgactgcggt gatattaccg gagaccttct gcgggacgag ccgggtcacg
      421 cggctgacgc ggagcgtccg ttgggcgaca aacaccagga cggggcacag gtacactatc
      481 ttgtcacccg gaggcgcgag ggactgcagg agcttcaggg agtggcgcag ctgcttcatc
      541 cccgtggccc gttgctcgcg tttgctggcg gtgtccccgg aagaaatata tttgcatgtc
      601 tttagttcta tgatgacaca aaccccgccc agcgtcttgt cattggcgaa gtcgaacacg
      661 cagatgcagt cggggcggcg cggtcccagg tccacttcgc atattaaggt gacgcgtgtg
      721 gcctcgaaca ccgagcgacc ctgcagcgac ccgcttaaaa gcttggcaat ccggtactgt
      781 tggtaaagcc accgctagcg ctcaccatga actccttctc cacaagcgcc ttcggtccag
      841 ttgccttctc cctgggcctg ctcctggtgt tgcctgctgc cttccctgcc ccagtgagcg
      901 gctggcggct gttcaagaag attagcggga gctccggtgg ctcgagcggt gcgatcgcca
      961 tggaataagt aaggaatcca catggcacag gttatcaaca cgtttgacgg ggttgcggat
     1021 tatcttcaga catatcataa gctacctgat aattacatta caaaatcaga agcacaagcc
     1081 ctcggctggg tggcatcaaa agggaacctt gcagacgtcg ctccggggaa aagcatcggc
     1141 ggagacatct tctcaaacag ggaaggcaaa ctcccgggca aaagcggacg aacatggcgt
     1201 gaagcggata ttaactatac atcaggcttc agaaattcag accggattct ttactcaagc
     1261 gactggctga tttacaaaac aacggaccat tatcagacct ttacaaaaat cagataatgt
     1321 ttaaacgaat tcgggctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag
     1381 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     1441 aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat
     1501 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1561 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1621 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     1681 aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg
     1741 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     1801 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     1861 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2041 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2101 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     2161 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttaatt
     2221 aactgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     2281 gaactaaacc caggaggcag atcatgattg aacaagatgg attgcacgca ggttctccgg
     2341 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2401 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2461 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2521 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2581 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2641 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2701 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2761 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2821 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2881 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2941 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     3001 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     3061 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     3121 gaccgaccaa gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt
     3181 cattacatct gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc
     3241 ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt
     3301 tctgcggact ggctttctac ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     3361 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4021 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4081 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4141 tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct gggatggtga
     4201 aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct gtaccctctg
     4261 gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag tttttcgggt
     4321 ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg
     4381 cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc catcgagata
     4441 aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcga
//
