LOCUS       sequence-447561-        7157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express gene of interest under EF-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7157)
  TITLE     lenti vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7157)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pCDH-EF1
FEATURES             Location/Qualifiers
     source          1..7157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(141..367)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(382..399)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(382..398)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(391..413)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(607..626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     726..748
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(786..804)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        872..976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(977..1045,1046..1837)
                     /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(1195..1214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2008..2596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2750..2767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2884..2905
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2920..2925,2926..2943,2944..2950)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2958..2974
                     /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)."
     primer_bind     2963..2985
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2982..2998
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2982..2998
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      3031..3166
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3206..3327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3217..3236)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3271..3290
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             3399..3632
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    3706..4294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3812..3823
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     4221..4241
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     promoter        complement(4331..5509)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4340..5278
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4363..4383)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    5607..5723
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(5990..6034)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    6219..6452
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    6945..7070
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             join(7117..7157,1..140)
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
       61 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
      121 tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac aatatctctg
      181 caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg ccttcctaat
      241 aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt tttctctcct
      301 tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag tattgcataa
      361 gactacacgc gtcagcttgg cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
      421 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
      481 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg
      541 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
      601 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
      661 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      721 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
      781 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
      841 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      961 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1021 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1081 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1141 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1201 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1261 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1321 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1381 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1441 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1501 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1561 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1621 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1681 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1741 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1801 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1981 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2041 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2101 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     2161 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2221 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2281 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2341 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2401 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2461 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2521 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2581 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2641 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2701 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2761 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2821 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2881 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     2941 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacat
     3001 gattacgaat ttggtctctg caaaagtcta ggcctccaaa aaagcctcct cactacttct
     3061 ggaatagctc agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc
     3121 catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg
     3181 gcgggatagc tagagccaga catgataaga tacattgatg agtttggaca aaccacaact
     3241 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     3301 accattataa gctgcaataa acaagttcct ctcactctct gatattcatt tctttgcaag
     3361 ttataaatac tgaataataa gatgacatga actactactg ctagagattt tccacactga
     3421 ctaaaagggt ctgagggatc tctagttacc agagtcacac aacagacggg cacacactac
     3481 ttgaagcact caaggcaagc tttattgagg cttaagcagt gggttcccta gttagccaga
     3541 gagctcccag gctcagatct ggtctaacca gagagaccca gtacaagcaa aaagcagatc
     3601 ttatcttcgt tgggagtgaa ttagcccttc cagtcccccc ttttctttta aaaagtggct
     3661 aagatctaca gctgccttgt aagtcattgg tcttaaaggt accaggcggg gaggcggccc
     3721 aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc cgcagagccg
     3781 gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg tagcagaagg
     3841 acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag gacgatgatt
     3901 tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc cagcagcggg
     3961 caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg aaagtcccgg
     4021 aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca gcaaacacag
     4081 tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga gacagcaacc
     4141 aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca tgatacaaag
     4201 gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga ataccagtca
     4261 atctttcaca aattttgtaa tccagaggtt gattgtcgac gaattcgcgg ccgcccgggg
     4321 atccaattcc tcacgacacc tgaaatggaa gaaaaaaact ttgaaccact gtctgaggct
     4381 tgagaatgaa ccaagatcca aactcaaaaa gggcaaattc caaggagaat tacatcaagt
     4441 gccaagctgg cctaacttca gtctccaccc actcagtgtg gggaaactcc atcgcataaa
     4501 acccctcccc ccaacctaaa gacgacgtac tccaaaagct cgagaactaa tcgaggtgcc
     4561 tggacggcgc ccggtactcc gtggagtcac atgaagcgac ggctgaggac ggaaaggccc
     4621 ttttcctttg tgtgggtgac tcacccgccc gctctcccga gcgccgcgtc ctccattttg
     4681 agctccctgc agcagggccg ggaagcggcc atctttccgc tcacgcaact ggtgccgacc
     4741 gggccagcct tgccgcccag ggcggggcga tacacggcgg cgcgaggcca ggcaccagag
     4801 caggccggcc agcttgagac tacccccgtc cgattctcgg tggccgcgct cgcaggcccc
     4861 gcctcgccga acatgtgcgc tgggacgcac gggccccgtc gccgcccgcg gccccaaaaa
     4921 ccgaaatacc agtgtgcaga tcttggcccg catttacaag actatcttgc cagaaaaaaa
     4981 gcgtcgcagc aggtcatcaa aaattttaaa tggctagaga cttatcgaaa gcagcgagac
     5041 aggcgcgaag gtgccaccag attcgcacgc ggcggcccca gcgcccaggc caggcctcaa
     5101 ctcaagcacg aggcgaaggg gctccttaag cgcaaggcct cgaactctcc cacccacttc
     5161 caacccgaag ctcgggatca agaatcacgt actgcagcca ggtggaagta attcaaggca
     5221 cgcaagggcc ataacccgta aagaggccag gcccgcggga accacacacg gcacttacct
     5281 gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact gcacttatat
     5341 acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca ctttcccagt
     5401 ttaccccgcg ccaccttctc taggcaccgg ttcaattgcc gacccctccc cccaacttct
     5461 cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcca attcccactc
     5521 ctttcaagac ctagaaggtc cattagctgc aaagattcct ctctgtttaa aactttatcc
     5581 atctttgcaa agctatcgat atcgataaaa ttttgaattt tgtaatttgt ttttgtaatt
     5641 ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca ctgtaccccc
     5701 caatcccccc ttttctttta aaagttaacc gataccgtcg agatccgttc actaatcgaa
     5761 tggatctgtc tctgtctctc tctccacctt cttcttctat tccttcgggc ctgtcgggtc
     5821 ccctcggggt tgggaggtgg gtctgaaacg ataatggtga atatccctgc ctaactctat
     5881 tcactataga aagtacagca aaaactattc ttaaacctac caagcctcct actatcatta
     5941 tgaataattt tatataccac agccaatttg ttatgttaaa ccaattccac aaacttgccc
     6001 atttatctaa ttccaataat tcttgttcat tcttttcttg ctggttttgc gattcttcaa
     6061 ttaaggagtg tattaagctt gtgtaattgt taatttctct gtcccactcc atccaggtcg
     6121 tgtgattcca aatctgttcc agagatttat tactccaact agcattccaa ggcacagcag
     6181 tggtgcaaat gagttttcca gagcaacccc aaatccccag gagctgttga tcctttaggt
     6241 atctttccac agccaggatt cttgcctgga gctgcttgat gccccagact gtgagttgca
     6301 acagatgctg ttgcgcctca atagccctca gcaaattgtt ctgctgctgc actataccag
     6361 acaataattg tctggcctgt accgtcagcg tcattgacgc tgcgcccata gtgcttcctg
     6421 ctgctcccaa gaacccaagg aacaaagctc ctattcccac tgctcttttt tctctctgca
     6481 ccactcttct ctttgccttg gtgggtgcta ctcctaatgg ttcaattttt actactttat
     6541 atttatataa ttcacttctc caattgtccc tcatatctcc tcctccaggt ctgaagatca
     6601 gtggccgctt gctgtgcggt ggtcttactt ttgttttgct cttcctctat cttgtctaaa
     6661 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
     6721 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
     6781 atttgtctac agccttctga tgtttctaac aggccaggat taactgcgaa tcgttctagc
     6841 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
     6901 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
     6961 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
     7021 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
     7081 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
     7141 aaaagggtct gagggat
//
