LOCUS       Exported                6614 bp ds-DNA     circular SYN 10-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6614)
  AUTHORS   jay
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 24, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6614
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        378..1556
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          609..1547
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1623..1641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1677..1839
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1840..2553
                     /codon_start=1
                     /label=NRL-203 (full length)
                     /translation="MALPPSPLAMEYVNDFDLMKFEVKREPSEGRPGPPTASLGSTPYS
                     SVPPSPTFSEPGMVGATEGTRPGLEELYWLATLQQQLGAGEALGLSPEEAMELLQGQGP
                     VPVDGPHGYYPGSPEETGAQHVQLAERFSDAALVSMSVRELNRQLRGCGRDEALRLKQR
                     RRTLKNRGYAQACRSKRLQQRRGLEAERARLAAQLDALRAEVARLARERDLYKARCDRL
                     TSSGPGSGDPSHLFL"
     polyA_signal    2627..2851
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2897..3325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3339..3669
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3520..3655
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3717..3764
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3783..4157
                     /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    4287..4408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4457..4473)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4481..4497
                     /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(4505..4535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4550..4571
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4859..5447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5618..6478)
                     /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"
     promoter        complement(6479..6583)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagcaaattg ggacattgat tattgactag gcttttgcaa aaagctttgc
      301 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      361 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      421 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      481 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      541 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      601 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      661 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      721 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      781 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
      841 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
      901 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
      961 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1021 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1081 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1141 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1201 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1261 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1321 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1381 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1441 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1501 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1561 ttagcttggt accgtcctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     1621 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttagcg
     1681 cagaggcttg gggcagccga gcggcagcca ggccccggcc cgggcctcgg ttccagaagg
     1741 gagaggagcc cgccaaggcg cgcaagagag cgggctgcct cgcagtccga gccggagagg
     1801 gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca tggccctgcc ccccagcccc
     1861 ctggccatgg aatatgtcaa tgactttgac ttgatgaagt ttgaggtaaa gcgggaaccc
     1921 tctgagggcc gacctggccc ccctacagcc tcactgggct ccacacctta cagctcagtg
     1981 cctccttcac ccaccttcag tgaaccaggc atggtggggg caaccgaggg cacccggcca
     2041 ggcctggagg agctgtactg gctggctacc ctgcagcagc agctgggggc tggggaggca
     2101 ttggggctga gtcctgaaga ggccatggag ctgctgcagg gtcagggccc agtccctgtt
     2161 gatgggcccc atggctacta cccagggagc ccagaggaga caggagccca gcacgtccag
     2221 ctggcagagc ggttttccga cgcggcgctg gtctcgatgt ctgtgcggga gctaaaccgg
     2281 cagctgcggg gctgcgggcg cgacgaggcg ctgcggctga agcagaggcg ccgcacgctg
     2341 aagaaccgcg gctacgcgca ggcctgtcgc tccaagcggc tgcagcagcg gcgcgggctg
     2401 gaggccgagc gcgcccgcct ggccgcccag ctggacgcgc tgcgggccga ggtggcccgc
     2461 ctggcccggg agcgcgatct ctacaaggct cgctgtgacc ggctaacctc gagcggcccc
     2521 gggtccgggg acccctccca cctcttcctc tgagccgttc agagcacctt gtggtggcgg
     2581 ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt
     2641 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2701 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2761 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2821 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2881 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2941 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3001 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     3061 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3121 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3181 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     3241 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     3301 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3361 agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3421 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3541 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3601 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3661 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca gcacgtgttg
     3721 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     3781 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     3841 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     3901 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     3961 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     4021 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     4081 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     4141 ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc ttctatgaaa
     4201 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4261 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4321 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4381 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4441 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4501 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4561 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4621 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5041 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5281 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6601 tgccacctga cgtc
//
