LOCUS       sequence-474762-        6006 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 6006)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6006)
  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..6006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..122
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12..31)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     66..85
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        367..724
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        367..558
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(390..410)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      575..710
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        739..786
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             818..1612
                     /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(872..891)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1482..1501
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1676..1724
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(1963..2551)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2043..2062)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2667..2950
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     promoter        3005..3756
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     primer_bind     complement(5952..5970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
       61 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      121 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
      181 ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga attttaacaa aatattaacg
      241 cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      301 tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa cttggttagg
      361 taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
      421 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      481 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      541 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      601 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
      661 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      721 caaaaagctt aattaactgt tgacaattaa tcatcggcat agtatatcgg catagtataa
      781 tacgacaagg tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca
      841 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
      901 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
      961 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     1021 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     1081 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     1141 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     1201 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     1261 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     1321 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     1381 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     1441 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     1501 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     1561 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     1621 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgatg gccgcaataa
     1681 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagcgataag
     1741 gatcctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     1801 gggtcagcac cgtttctgcg gactggcttt ctacccggta tcagctcact caaaggcggt
     1861 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1921 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1981 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2041 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2101 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2161 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2221 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2281 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2341 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2401 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2461 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2521 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     2581 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2641 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     2701 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     2761 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     2821 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     2881 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     2941 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     3001 tctaaatgag tcttcggacc tcgcgggggc cgcttaagcg gtggttaggg tttgtctgac
     3061 gcggggggag ggggaaggaa cgaaacactc tcattcggag gcggctcggg gtttggtctt
     3121 ggtggccacg ggcacgcaga agagcgccgc gatcctctta agcacccccc cgccctccgt
     3181 ggaggcgggg gtttggtcgg cgggtggtaa ctggcgggcc gctgactcgg gcgggtcgcg
     3241 cgccccagag tgtgaccttt tcggtctgct cgcagacccc cgggcggcgc cgccgcggcg
     3301 gcgacgggct cgctgggtcc taggctccat ggggaccgta tacgtggaca ggctctggag
     3361 catccgcacg actgcggtga tattaccgga gaccttctgc gggacgagcc gggtcacgcg
     3421 gctgacgcgg agcgtccgtt gggcgacaaa caccaggacg gggcacaggt acactatctt
     3481 gtcacccgga ggcgcgaggg actgcaggag cttcagggag tggcgcagct gcttcatccc
     3541 cgtggcccgt tgctcgcgtt tgctggcggt gtccccggaa gaaatatatt tgcatgtctt
     3601 tagttctatg atgacacaaa ccccgcccag cgtcttgtca ttggcgaagt cgaacacgca
     3661 gatgcagtcg gggcggcgcg gtcccaggtc cacttcgcat attaaggtga cgcgtgtggc
     3721 ctcgaacacc gagcgaccct gcagcgaccc gcttaaaagc ttggcaatcc ggtactgttg
     3781 gtaaagccac cgctagcgct caccatggtg agcggctggc ggctgttcaa gaagattagc
     3841 gggagctccg gtggctcgag cggtgcgatc gccatggagc ggaaagtgct tgcgctccag
     3901 gcccgaaaga aaaggaccaa ggccaagaag gacaaagccc aaaggaaatc tgaaactcag
     3961 caccgaggct ctgctcccca ctctgagagt gatctaccag agcaggaaga ggagattctg
     4021 ggatctgatg atgatgagca agaagatcct aatgattatt gtaaaggagg ttatcatctt
     4081 gtgaaaattg gagatctatt caatgggaga taccatgtga tccgaaagtt aggctgggga
     4141 cacttttcaa cagtatggtt atcatgggat attcagggga agaaatttgt ggcaatgaaa
     4201 gtagttaaaa gtgctgaaca ttacactgaa acagcactag atgaaatccg gttgctgaag
     4261 tcagttcgca attcagaccc taatgatcca aatagagaaa tggttgttca actactagat
     4321 gactttaaaa tatcaggagt taatggaaca catatctgca tggtatttga agttttgggg
     4381 catcatctgc tcaagtggat catcaaatcc aattatcagg ggcttccact gccttgtgtc
     4441 aaaaaaatta ttcagcaagt gttacagggt cttgattatt tacataccaa gtgccgtatc
     4501 atccacactg acattaaacc agagaacatc ttattgtcag tgaatgagca gtacattcgg
     4561 aggctggctg cagaagcaac agaatggcag cgatctggag ctcctccgcc ttccggatct
     4621 gcagtcagta ctgctcccca gcctaaacca gctgacaaaa tgtcaaagaa taagaagaag
     4681 aaattgaaga agaagcagaa gcgccaggca gaattactag agaagcgaat gcaggaaatt
     4741 gaggaaatgg agaaagagtc gggccctggg caaaaaagac caaacaagca agaagaatca
     4801 gagagtcctg ttgaaagacc cttgaaagag aacccaccta ataaaatgac ccaagaaaaa
     4861 cttgaagagt caagtaccat tggccaggat caaacgctta tggaacgtga tacagagggt
     4921 ggtgcagcag aaattaattg caatggagtg attgaagtca ttaattatac tcagaacagt
     4981 aataatgaaa cattgagaca taaagaggat ctacataatg ctaatgactg tgatgtccaa
     5041 aatttgaatc aggaatctag tttcctaagc tcccaaaatg gagacagcag cacatctcaa
     5101 gaaacagact cttgtacacc tataacatct gaggtgtcag acaccatggt gtgccagtct
     5161 tcctcaactg taggtcagtc attcagtgaa caacacatta gccaacttca agaaagcatt
     5221 cgggcagaga taccctgtga agatgaacaa gagcaagaac ataacggacc actggacaac
     5281 aaaggaaaat ccacggctgg aaattttctt gttaatcccc ttgagccaaa aaatgcagaa
     5341 aagctcaagg tgaagattgc tgaccttgga aatgcttgtt gggtgcacaa acatttcact
     5401 gaagatattc aaacaaggca atatcgttcc ttggaagttc taatcggatc tggctataat
     5461 acccctgctg acatttggag cacggcatgc atggcctttg aactggccac aggtgactat
     5521 ttgtttgaac ctcattcagg ggaagagtac actcgagatg aagatcacat tgcattgatc
     5581 atagaacttc tggggaaggt gcctcgcaag ctcattgtgg caggaaaata ttccaaggaa
     5641 tttttcacca aaaaaggtga cctgaaacat atcacgaagc tgaaaccttg gggccttttt
     5701 gaggttctag tggagaagta tgagtggtcg caggaagagg cagctggctt cacagatttc
     5761 ttactgccca tgttggagct gatccctgag aagagagcca ctgccgccga gtgtctccgg
     5821 cacccttggc ttaactccgt ttaaacgaat tcgggctcgg tacccgggga tcctctagag
     5881 tcgacctgca ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     5941 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg cttcgagcag
     6001 acatga
//
