LOCUS       sequence-243321-        6775 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 6775)
  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 6775)
  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-10A1
FEATURES             Location/Qualifiers
     source          1..6775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          316..888
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     RBS             1933..1941
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     primer_bind     complement(3003..3022)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3068..3123
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3122..3141)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3482..3498)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3482..3498)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3495..3517)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3506..3522
                     /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(3530..3536,3537..3554,3555..3560))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3575..3596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3713..3730)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3884..4472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3964..3983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4643..5434,5435..5503))
                     /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     5266..5285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5504..5608)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5634..6089)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5771..5792)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5983..6002
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6216..6238
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6230..6247
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6231..6247
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6257..6276
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6257..6275
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        6396..6775
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gtcgaccgat
      301 cctgagaact tcagggtgag tttggggacc cttgattgtt ctttcttttt cgctattgta
      361 aaattcatgt tatatggagg gggcaaagtt ttcagggtgt tgtttagaat gggaagatgt
      421 cccttgtatc accatggacc ctcatgataa ttttgtttct ttcactttct actctgttga
      481 caaccattgt ctcctcttat tttcttttca ttttctgtaa ctttttcgtt aaactttagc
      541 ttgcatttgt aacgaatttt taaattcact tttgtttatt tgtcagattg taagtacttt
      601 ctctaatcac ttttttttca aggcaatcag ggtatattat attgtacttc agcacagttt
      661 tagagaacaa ttgttataat taaatgataa ggtagaatat ttctgcatat aaattctggc
      721 tggcgtggaa atattcttat tggtagaaac aactacaccc tggtcatcat cctgcctttc
      781 tctttatggt tacaatgata tacactgttt gagatgagga taaaatactc tgagtccaaa
      841 ccgggcccct ctgctaacca tgttcatgcc ttcttctctt tcctacagct cctgggcaac
      901 gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tcctcgacgg atctgggccg
      961 caccatggcg cgttcaacgc tctcaaaacc ccttaaagat aagattaacc cgtggaagtc
     1021 cttaatggtc atgggggtcc tattaagagt agggatggca gagagccccc atcaggtctt
     1081 taatgtaacc tggagagtca ccaacctgat gaccgggcgt accgccaatg ccacctccct
     1141 tttaggaact gtacaagatg ccttcccaag attatatttt gatctatgtg acctggtcgg
     1201 agaagagtgg gacccttcag accaggaacc atatgtcggg tatggctgca aataccccgg
     1261 agggagaaag cggacccgga cttttgactt ttacgtgtgc cctgggcata ccgtaaaatc
     1321 ggggtgtggg gggccaagag aaggctactg tggtgaatgg ggttgtgaaa ccaccggaca
     1381 ggcttactgg aagcccacat catcatggga cctaatctcc cttaagcgcg gtaacacccc
     1441 ctgggacacg ggatgctcca aaatggcttg tggcccctgc tatgacctct ccaaggtatc
     1501 caattccttc caaggggcta ctcgaggggg cagatgcaac cctctagtcc tagaattcac
     1561 tgatgcagga aaaaaggcta attgggacgg gcccaaatcg tggggactga gactgtaccg
     1621 gacaggaaca gatcctatta ccatgttctc cctgacccgc caggtcctca atatagggcc
     1681 ccgcatcccc attgggccta atcccgtgat cactggtcaa ctacccccct cccgacccgt
     1741 gcagatcagg ctccccaggc ctcctcagcc tcctcctaca ggcgcagcct ctatagtccc
     1801 tgagactgcc ccaccttctc aacaacctgg gacgggagac aggctgctaa acctggtaga
     1861 aggagcctat cgggcgctta acctcaccaa tcccgacaag acccaagaat gttggctgtg
     1921 cttagtgtcg ggacctcctt attacgaagg agtagcggtc gtgggcactt ataccaatca
     1981 ttctaccgcc ccggccagct gtacggccac ttcccaacat aagcttaccc tatctgaagt
     2041 gacaggacag ggcctatgca tgggagcagt acctaaaact caccaggcct tatgtaacac
     2101 cacccaaagt gccggctcag gatcctacta ccttgcagca cccgctggaa caatgtgggc
     2161 ttgtagcact ggattgactc cctgcttgtc caccacgatg ctcaatctaa ccacagacta
     2221 ttgtgtatta gttgagctct ggcccagaat aatttaccac tcccccgatt atatgtatgg
     2281 tcagcttgaa cagcgtacca aatataagag ggagccagta tcgttgaccc tggcccttct
     2341 gctaggagga ttaaccatgg gagggattgc agctggaata gggacgggga ccactgccct
     2401 aatcaaaacc cagcagtttg agcagcttca cgccgctatc cagacagacc tcaacgaagt
     2461 cgaaaaatca attaccaacc tagaaaagtc actgacctcg ttgtctgaag tagtcctaca
     2521 gaaccgaaga ggcctagatt tgctcttcct aaaagaggga ggtctctgcg cagccctaaa
     2581 agaagaatgt tgtttttatg cagaccacac gggactagtg agagacagca tggccaaact
     2641 aagggaaagg cttaatcaga gacaaaaact atttgagtca ggccaaggtt ggttcgaagg
     2701 gctgtttaat agatccccct ggtttaccac cttaatctcc accatcatgg gacctctaat
     2761 agtactctta ctgatcttac tctttggacc ctgcattctc aatcgattag tccaatttgt
     2821 taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc accagctgaa
     2881 gcccatagag tacgagccat gataaccaga tccgtcgagg aattcactcc tcaggtgcag
     2941 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     3001 gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact
     3061 tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc
     3121 tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt
     3181 tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga
     3241 ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg
     3301 acttgaggtt agattttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     3361 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     3421 catagctgtc cctcttctct tatggagatc cctcgacgga tcggccgcaa ttcgtaatca
     3481 tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3541 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3601 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3661 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3721 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3781 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3841 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4321 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4921 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     5641 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     5701 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     5761 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     5821 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     5881 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     5941 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6001 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6061 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     6121 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6181 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     6241 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctcc accgcggtgg
     6301 cggccgctct agagagcttg gcccattgca tacgttgtat ccatatcata atatgtacat
     6361 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
     6421 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6481 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6541 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6601 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6661 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6721 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
