LOCUS       Exported                5900 bp ds-DNA     circular SYN 23-JUL-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5900)
  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..5900
                     /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"
     polyA_signal    1913..2137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2183..2611
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2625..2955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2806..2941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3003..3050
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3069..3443
                     /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    3573..3694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3743..3759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3767..3783
                     /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(3791..3821)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3836..3857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4904..5764)
                     /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(5765..5869)
                     /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 tccgaaaccg ccgttcagag caccttgtgg
     1861 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1921 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1981 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     2041 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     2101 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2161 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2221 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2281 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     2341 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2401 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2461 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2521 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     2581 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2641 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     2701 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     2761 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     2881 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2941 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     3001 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3061 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
     3121 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
     3181 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
     3241 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
     3301 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
     3361 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
     3421 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
     3481 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3541 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3601 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3661 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3721 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3781 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3841 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3901 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3961 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4801 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4861 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4921 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4981 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     5041 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     5101 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5161 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5221 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     5281 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5341 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5401 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5461 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5521 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5581 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5641 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5701 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5761 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5821 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5881 gaaaagtgcc acctgacgtc
//
