LOCUS       Exported                6143 bp ds-DNA     circular SYN 08-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6143)
  AUTHORS   Heegwang Roh
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 6, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6143
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1694..1740
                     /label=HR572_fwd_EcoRI
     regulatory      1726..1735
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1735..1776
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    1813..3624
                     /label=LATeNT
     primer_bind     complement(3601..3658)
                     /label=HR572_rev_HindIII
     primer_bind     complement(3646..3662)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3670..3686
                     /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(3694..3724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3739..3760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3819..4014
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3865..4000
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4020..4154
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4393..4981)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5152..6012)
                     /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(6013..6117)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attccgccac catgggcaag
     1741 cccatcccca accccctgct gggcctggac agcaccaagg gctcgggctc gacctcgggc
     1801 tcgggcaccg gtccgatcac catcaacaac ttccgttact ccgacccggt taacaacgac
     1861 accatcatca tgatggaacc gccgtactgc aaaggtctag acatctacta cagagccttc
     1921 aaaatcaccg accgtatctg gatcgttccg gaacgttacg aatttggtac caaaccggaa
     1981 gacttcaacc cgccgtcctc cctgatcgaa ggtgcttccg aatactacga cccgaactac
     2041 ctgcgtaccg actccgacaa agaccgtttc ctgcagacca tggttaaact gttcaaccgt
     2101 atcaaaaaca acgttgctgg tgaagctctg ctggacaaaa tcatcaacgc tatcccgtac
     2161 ctgggtaact cctactccct gctggacaaa ttcgacacca actccaactc cgtttccttc
     2221 aacctgctgg aacaggatcc gtccggtgct accaccaaat ccgctatgct gaccaacctg
     2281 atcatcttcg gtccgggtcc ggttctgaac aaaaacgaag ttcgtggtat cgttctgcgt
     2341 gttgacaaca aaaactactt cccgtgccgt gacggtttcg gttccatcat gcagatggct
     2401 ttctgcccgg aatacgtacc gaccttcgac aacgttatcg aaaacatcac ctccctgacc
     2461 atcggtaaat ccaaatactt ccaggacccg gctctgctgc tgatgcacga actgatccac
     2521 gttctgcacg gtctgtacgg catgcaggtt tcctcccacg aaatcatccc gtccaaacag
     2581 gaaatctaca tgcagcacac ctacccgatc tccgctgaag agctcttcac cttcggtggt
     2641 caggacgcta acctgatctc catcgatatc aaaaacgacc tgtacgaaaa aaccctgaac
     2701 gactacaaag ctatcgctaa caaactgtcc caggttacct cctgcaacga cccgaacatc
     2761 gacatcgact cctacaaaca gatctatcag cagaagtatc agtttgataa ggattccaac
     2821 ggccagtata tagtgaacga ggataagttt cagatactgt ataactccat aatgtatggc
     2881 gggagcggca gcggggggtc cggtagcggg gagtttcggg caaccacact ggaacggatc
     2941 gagaaaagct tcgtgattac tgatccgaga ctgcctgaca acccaatcat ttttgtgagc
     3001 aattccttcc tgcagctgac agaatattct cgggaagaga tcctggggcg caattgccgt
     3061 tttctgcagg gacccgagac agaccgtgcc actgttcgga aaatcagaga tgctattgac
     3121 aaccagactg aagtgaccgt tcagctgatc aattatacca agagcggcaa gaagttctgg
     3181 aacgtgttcc acctgcagcc gatgcgcgat tataagggcg acgtccagta cttcattggc
     3241 gtgcagctgg atggcaccga acgccttcat ggcgccgctg agcgtgaggc ggtctgcctg
     3301 gtcaaaaaga cagcctttca gatttttact gagatagagc tgggcaagaa gtttaacata
     3361 aagactcgct taagctattt ttccatgaac cacgatcctg tgaagatacc taacctgctg
     3421 gatgatacta tatataacga tactgagggc tttaacatag agtccaagga tctgaagtcc
     3481 gagtataagg gccagaacat gagggtgaac actaacgcct ttaggaacgt cgacggttcc
     3541 ggtctggttt ccaaactgat cggtctgtgc aaaaaaatca tcccgccgac caacatccgt
     3601 gaaaacctgt acaaccgtac cgcttaaaag cttggcgtaa tcatggtcat agctgtttcc
     3661 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     3721 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     3841 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3901 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3961 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     4021 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4081 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4141 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4201 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4261 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4321 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4381 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4441 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4501 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4561 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     4621 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4681 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4741 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4801 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4861 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4921 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4981 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5041 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5101 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5161 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5221 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5281 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5341 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5401 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5461 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5521 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5581 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5641 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5701 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5761 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5821 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5881 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5941 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6001 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6061 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6121 atttccccga aaagtgccac ctg
//
