LOCUS       sequence-148124-        6483 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6483)
  TITLE     Cavb3
REFERENCE   2  (bases 1 to 6483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Cavb3
FEATURES             Location/Qualifiers
     source          1..6483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2257..2274)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2263..2487
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2533..2961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2620..2639)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2830..2851
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2970..2990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2975..3304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3155..3290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3217..3236
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3352..3399
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3418..3792
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3922..4043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3959..3978)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4013..4032
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4092..4108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4092..4108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4105..4127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4116..4132
                     /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(4140..4146,4147..4164,4165..4170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4185..4206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4323..4340)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4494..5082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4574..4593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5253..6044,6045..6113))
                     /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     5876..5895
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6114..6218)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6293..6312)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc cccggactcc cccatgtatg
      721 acgactccta cgtgcccggg tttgaggact cggaggcggg ttcagccgac tcctacacca
      781 gccgcccctc tctggactca gacgtttccc tggaggagga ccgggagagt gcccggcgag
      841 aagtggagag tcaggctcag cagcagctgg aaagagccaa gcacaaacct gtggcatttg
      901 ctgtgaggac caatgtcagc tactgtggag ttctggatga ggaatgccca gtccagggct
      961 ctggagtcaa cttcgaggcc aaagattttc tgcacattaa agagaagtac agcaatgact
     1021 ggtggatcgg gaggctagtg aaagaaggtg gcgatattgc cttcatcccc agcccccaac
     1081 gcctggagag catccggctc aaacaggaac agaaggccag gagatccggg aacccttcca
     1141 gcctgagtga cattggcaac cgacgttccc ctcctccatc tctagccaag cagaagcaaa
     1201 agcaggcgga acatgtcccc ccgtatgatg tggtgccctc catgcggcct gtggtgctgg
     1261 tgggaccctc tctgaaaggt tatgaggtca cagacatgat gcagaaggct ctcttcgact
     1321 tccttaaaca caggtttgat ggcaggatct ccatcacccg cgtcacggct gacctctcac
     1381 tggccaagcg ctctgtgctc aacaatcctg gcaagaggac catcatcgag cgctcttctg
     1441 cccgctccag cattgctgag gtgcagagtg agattgagcg catattcgag ctggccaaat
     1501 ccctgcagct agtggtgttg gatgctgaca ccatcaacca cccagcacag ctagccaaga
     1561 cctcactggc ccccatcatc gtcttcgtca aagtgtcctc gccaaaggta ctgcagcgac
     1621 tgatccgctc cagggggaag tcccagatga agcacctcac tgtacagatg atggcatatg
     1681 ataagctggt tcagtgccca cctgagtcat ttgatgtgat tctggatgag aaccagctgg
     1741 acgacgcctg tgagcaccta gctgaatacc tagaggttta ctggcgcgct acccaccacc
     1801 cagcaccggg ccccgggatg ctgggtccgc ccagtgccat ccctggactt cagaaccagc
     1861 agctgctggg ggagcgaggt gaggagcatt cacccctgga gcgggacagt ttgatgccct
     1921 cggatgaggc cagtgagagc tcccgccagg cctggaccgg atcttcacag cgcagctccc
     1981 gccatctgga ggaggactat gcagatgcct accaggacct gtaccagcct caccgtcaac
     2041 acacctcggg gctacccagt gctaacgggc atgaccccca agaccggctc ctagcccagg
     2101 actcggagca tgaccacaat gaccggaact ggcagcgtaa ccggccttgg cctaaggaca
     2161 gctactgacc acctcctgcc ctacccgaat tctgcagata tccagcacag tggcggccgc
     2221 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     2281 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     2341 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     2401 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     2461 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     2521 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2581 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2641 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2701 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2761 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2821 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2881 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2941 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3241 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3301 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     3361 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     3421 gccaagttga ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag
     3481 ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg
     3541 gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac
     3601 accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc
     3661 gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg
     3721 tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag
     3781 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     3841 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3901 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3961 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     4021 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     4081 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4141 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4201 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4261 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     4321 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4381 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4441 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4501 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4561 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4621 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4681 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4741 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4801 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4861 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4921 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4981 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5041 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5101 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5161 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5221 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5281 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5341 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5401 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5461 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5521 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5581 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5641 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5701 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5761 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5821 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5881 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5941 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6001 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6061 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6121 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     6181 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     6241 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     6301 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     6361 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     6421 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     6481 gtt
//
