LOCUS       sequence-8353-ex        6763 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6763)
  AUTHORS   Sena-Esteves M, Tebbets JC, Steffens S, Crombleholme T, Flake AW
  TITLE     Optimized large-scale production of high titer lentivirus vector 
            pseudotypes.
  JOURNAL   J Virol Methods. 2004 Dec 15. 122(2):131-9.
  PUBMED    15542136
REFERENCE   2  (bases 1 to 6763)
  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: "J Virol 
            Methods. 2004 Dec 15. 122(2):131-9."
COMMENT     Sequence Label: pHCMV-EcoEnv
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        477..680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     631..651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     677..701
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          792..1364
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3467..3486)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3532..3587
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3586..3605)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3945..3961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3945..3961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3958..3980)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3969..3985
                     /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)."
     promoter        complement(join(3993..3999,4000..4017,4018..4023))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4038..4059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4176..4193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4347..4935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4427..4446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5404..5783)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     5730..5749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5968..6072)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6098..6553)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6235..6256)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6447..6466
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6680..6702
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6694..6711
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6695..6711
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6721..6740
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6721..6739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcggccgctc tagagagctt ggcccattgc atacgttgta tccatatcat aatatgtaca
       61 tttatattgg ctcatgtcca acattaccgc catgttgaca ttgattattg actagttatt
      121 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      181 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      241 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      301 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      361 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      421 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      481 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      541 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      601 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      661 aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac
      721 gctgttttga cctccataga agacaccggg accgatccag cctccggtcg accgatcctg
      781 agaacttcag ggtgagtttg gggacccttg attgttcttt ctttttcgct attgtaaaat
      841 tcatgttata tggagggggc aaagttttca gggtgttgtt tagaatggga agatgtccct
      901 tgtatcacca tggaccctca tgataatttt gtttctttca ctttctactc tgttgacaac
      961 cattgtctcc tcttattttc ttttcatttt ctgtaacttt ttcgttaaac tttagcttgc
     1021 atttgtaacg aatttttaaa ttcacttttg tttatttgtc agattgtaag tactttctct
     1081 aatcactttt ttttcaaggc aatcagggta tattatattg tacttcagca cagttttaga
     1141 gaacaattgt tataattaaa tgataaggta gaatatttct gcatataaat tctggctggc
     1201 gtggaaatat tcttattggt agaaacaact acaccctggt catcatcctg cctttctctt
     1261 tatggttaca atgatataca ctgtttgaga tgaggataaa atactctgag tccaaaccgg
     1321 gcccctctgc taaccatgtt catgccttct tctctttcct acagctcctg ggcaacgtgc
     1381 tggttgttgt gctgtctcat cattttggca aagaattccg cgggcggccg ctctagagga
     1441 tccgctagac tgacatggcg cgttcaacgc tctcaaaacc ccttaaaaat aaggttaacc
     1501 cgcgaggccc cctaatcccc ttaattcttc tgatgctcag aggggtcagt actgcttcgc
     1561 ccggctccag tcctcatcaa gtctataata tcacctggga ggtaaccaat ggagatcggg
     1621 agacggtatg ggcaacttct ggcaaccacc ctctgtggac ctggtggcct gaccttaccc
     1681 cagatttatg tatgttagcc caccatggac catcttattg ggggctagaa tatcaatccc
     1741 ctttttcttc tcccccgggg cccccttgtt gctcaggggg cagcagccca ggctgttcca
     1801 gagactgcga agaaccttta acctccctca cccctcggtg caacactgcc tggaacagac
     1861 tcaagctaga ccagacaact cataaatcaa atgagggatt ttatgtttgc cccgggcccc
     1921 accgcccccg agaatccaag tcatgtgggg gtccagactc cttctactgt gcctattggg
     1981 gctgtgagac aaccggtaga gcttactgga agccctcctc atcatgggat ttcatcacag
     2041 taaacaacaa tctcacctct gaccaggctg tccaggtatg caaagataat aagtggtgca
     2101 accccttagt tattcggttt acagacgccg ggagacgggt tacttcctgg accacaggac
     2161 attactgggg cttacgtttg tatgtctccg gacaagatcc agggcttaca tttgggatcc
     2221 gactcagata ccaaaatcta ggaccccgcg tcccaatagg gccaaacccc gttctggcag
     2281 accaacagcc actctccaag cccaaacctg ttaagtcgcc ttcagtcacc aaaccaccca
     2341 gtgggactcc tctctcccct acccaacttc caccggcggg aacggaaaat aggctgctaa
     2401 acttagtaga cggagcctac caagccctca acctcaccag tcctgacaaa acccaagagt
     2461 gctggttgtg tctagtagcg ggacccccct actacgaagg ggttgccgtc ctgggtacct
     2521 actccaacca tacctctgct ccagccaact gctccgtggc ctcccaacac aagttgaccc
     2581 tgtccgaagt gaccggacag ggactctgca taggagcagt tcccaaaaca catcaggccc
     2641 tatgtaatac cacccagaca agcagtcgag ggtcctatta tctagttgcc cctacaggta
     2701 ccatgtgggc ttgtagtacc gggcttactc catgcatctc caccaccata ctgaacctta
     2761 ccactgatta ttgtgttctt gtcgaactct ggccaagagt cacctatcat tcccccagct
     2821 atgtttacgg cctgtttgag agatccaacc gacacaaaag agaaccggtg tcgttaaccc
     2881 tggccctatt attgggtgga ctaaccatgg ggggaattgc cgctggaata ggaacaggga
     2941 ctactgctct aatggccact cagcaattcc agcagctcca agccgcagta caggatgatc
     3001 tcagggaggt tgaaaaatca atctctaacc tagaaaagtc tctcacttcc ctgtctgaag
     3061 ttgtcctaca gaatcgaagg ggcctagact tgttatttct aaaagaagga gggctgtgtg
     3121 ctgctctaaa agaagaatgt tgcttctatg cggaccacac aggactagtg agagacagca
     3181 tggccaaatt gagagagagg cttaatcaga gacagaaact gtttgagtca actcaaggat
     3241 ggtttgaggg actgtttaac agatcccctt ggtttaccac cttgatatct accattatgg
     3301 gacccctcat tgtactccta atgattttgc tcttcggacc ctgcattctt aatcgattag
     3361 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     3421 accagctgaa gcctatagag tacgagccat agctcgaggg cccagatctt tttccctctg
     3481 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3541 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3601 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3661 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3721 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3781 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3841 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3901 tcttatggag atccctcgac ggatcggccg caattcgtaa tcatgtcata gctgtttcct
     3961 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4021 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4081 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4141 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4201 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4261 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4321 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4381 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4441 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4501 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4561 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4621 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4681 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4741 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4801 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4861 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4921 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4981 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5041 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5101 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5161 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5221 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5281 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5341 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5401 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5461 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5521 aaagcgggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     5761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5941 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6001 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6061 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
     6121 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6181 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     6241 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6301 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     6361 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6421 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6481 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6541 ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     6601 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     6661 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg
     6721 taatacgact cactataggg cgaattggag ctccaccgcg gtg
//
